TW200837075A - Antiviral peptides - Google Patents

Abstract

The present application is directed to antiviral peptides, methods of using these peptides to prevent or inhibit infections by a human immunodeficiency virus or a virus from the Flaviviridae family, and pharmaceutical compositions and combinations, as well as articles of manufacture comprising these peptides.

Description

200837075 IX. Description of the invention: [Technical field to which the invention pertains 1 Government sponsorship The present invention was funded by the US Government 5 under the National Health Service sponsorship number CA108304. The U.S. Government has certain rights in the invention. FIELD OF THE INVENTION The present invention relates to antiviral peptides. [Prior Art 2 Background of the Invention Viral diseases are difficult to treat because the virus enters mammalian cells and performs a number of viral functions in the cells, including transcription and translation of viral proteins, and replication of viral genomes. Thus, when a viral infection is carried out, the virus is not only protected from the immune system of the host animal, but is also protected from exposure to drugs administered to the host animal. There are already a few effective antiviral agents available, but most antiviral agents are only useful for a small group of viruses. Therefore, there is a need for an agent for treating and preventing a viral infection. I: SUMMARY OF THE INVENTION j SUMMARY OF THE INVENTION The present invention relates to peptides which prevent or treat infection by members of the human immunodeficiency virus (HIV) and Flaviviridae family. The invention also relates to peptides which are capable of deactivating such viruses before they enter the cell. The present invention is based on the Faxian dry antiviral peptide, and the plurality of peptides thereof have an amphipathic α-helical structure and a specific amino acid composition. Unexpectedly, a variety of such peptides can be deactivated in a solution free of virus, thereby preventing viral infection of mammalian cells. At the same time, surprisingly, the peptide is derived from and/or associated with hepatitis C polyprotein, but can deactivate the flavivirus, paramyxovirus, and human immunodeficiency virus (HIV). 5 The present invention relates to the discovery that an amphipathic a-helical peptide (SEQ ID NO: 43) derived from the membrane anchoring functional site of the hepatitis C virus (HCV) NS5A protein is virucidal to HCV, ie, The amphiphilic helix peptide in the tube deactivates HCV. The peptide can prevent repeated infection of HCV and inhibit infection by 10 by purifying extracellular infectious particles and intracellular infectious particles. In addition to HCV, the peptide inhibits selected flaviviruses, paramyxoviruses, and HIV infections. The peptide is also non-toxic in vitro and in vivo at doses at least 100 times greater than the dose required for antiviral activity. Its amphipathic 〇: _ helical structure is required, but for virucidal activity is insufficient, viricidal activity is based on its amino acid composition, rather than on its primary sequence or its 15 pairs of palmity. In one aspect, the present invention provides an isolated peptide having from 14 to 50 guanidine-amino acids or L-amino acids, comprising an amino acid sequence:

XaarXaa2-Xaa3-WL-Xaa6-Xaa7-Xaa8-W-Xaai〇-W-Xaa12-Xaa13-Xaai4-Xaai5-Xaai6-Xaa17-Xaai8-Xaa19-Xaa2〇-Xaa2rXaa22 (SEQ ID NO: 163), 20 of which Xaai is or is absent in the presence of serine (S); Xaa2 is glycine (G) or absent; Xaa3 is serine (S), aspartic acid (D) or threonine (T); Is tryptophan; L is leucine; Xaa6 is arginine (R), aspartic acid (D), lysine (K), tryptophan (W), or glutamic acid (E); Xaa7 Is aspartic acid (D), arginine (R), face acid (E), alanine (A), isoleucine (1), or amine 200837075 acid (Κ), Xaas is isoleucine (1) Or proline (v); also known as aspartic acid (9), arginine (R), glutamic acid (6) or lysine (κ); χ·ΐ2 is isoleucine (1) or leucine ( V) ; Xaan is cysteine (〇, face acid (6), leucine (L), serine (S) or arginine (r); xaai4 is glutamic acid (6), lysine 5, Aspartic acid (D), threonine (T), histidine (H), serine (8) or arginine (R); Xaa 5 is valine (V), alanine (Α) or Serine (8); xaai6 is leucine (L) or lysine (v); Xaa is 4 (serine), sulphate (τ), leucine (L) or is absent; xaa!8 is aspartic acid (D), arginine (R), glutamic acid (E), lysine (κ) or is absent; xaai9 is phenylalanine Or 10 is absent; Xaa2 is lysine (K), glutamic acid (E), arginine (R), aspartic acid (D) or is absent, Xaa2i is sulphate (T) or Is absent; and Xaa22 is a tryptophanic acid (W) or is absent; and wherein the amino acid sequence of the formula I is not: SWLRDIWDWICEVLSDFK (SEQ ID NO: 43); SWLRDIWDWICEVLSDF (SEQ ID NO: 95)? 15 SWLRDIWDWICEVLSD (SEQ ID NO: 94)? SWLRDIWDWICEVLS (SEQ ID NO: 93), or SWLRDIWDWICEVL (SEQ ID NO: 92) In several embodiments, the isolated peptide of the present invention comprises an amino acid sequence of formula II: 20 Xaai -Xaa2_S_ W_L_Xa3^-Xaa7_I_W-Xaai 〇W_I_C-

Xaa^-VL-Xaan-XaawX(10)i9-X(10)2(rXaa2i_Xaa22(SEQ ID NO: 164), wherein Xaa!, Xaa2, Xaa2i and Xaa22 are absent, S is serine; W is tryptophan; L is white amine Acid; Xaa6 is arginine (R), aspartic acid (D), glutamic acid (E) or lysine (K); Xaa7 is arginine (R), aspartic acid (D) or from 7 200837075 Aminic acid (κ); i is isoleucine; w is tryptophan; 1 is aspartic acid (D), arginine (R), or lysine (K); c is half Cysteine; Xaai4 is lysine (K), arginine (R), face acid (E) or aspartic acid (D); V is valine (V); Xaan is serine (S) Or is absent; Xaais is aspartic acid (D), arginine 5, lysine (K) or absent; Xaa^ is phenylalanine (F) or is absent; and Xaa2 is Amino acid, glutamic acid (6), aspartic acid, arginine (R) or absent. In several embodiments, the isolated peptide of the invention comprises the amino acid sequence of formula m: 10 XaarXaa2-SW -LR.Xaa7-IW-Xaa10.WI-C.

Xaa14_V-L-Xaa17_Xaa18_Xaa19_Xaa2 (rXaa2rXaa22 (SEQ ID NO: 178), wherein: Xaa, χ^Αχ^22 is absent; s is serine; W is tryptophan; L is leucine; R is Arginine; i is isoleucine; C is cysteine; V is valine; xaa? is aspartic acid (D), arginine (R) 15 or lysine (K); XaaiG Is arginine (R) or lysine (K); Xaai4 is lysine (K), glutamic acid (E) or aspartic acid (D" Xaai7 is seric acid (s) or is absent; Xaals is arginine (R), lysine (κ) or is absent; Xaai9 is phenylalanine (F) or is absent; and Xaa2 is lysine (K) or is absent. In several embodiments, the peptide of the present invention comprises an amino acid sequence of the formula: 2〇Xaai -Xaa2-SWL-Xaa6-Xaa7-IW-Xaai〇-WIC-

Xaai4-VL-Xaai7-Xaai8-Xaai9-Xaa2〇-Xaa2rXaa22 (SEQ ID NO: 184) wherein Xaai, Xaa], Xaa2i and Xaa22 are absent; s is serine; W is tryptophan; L is white Aminic acid; I is isoleucine; C is cysteine; 200837075 V is lysine, and Xaa6, X(10)7, X(10)ίο and X(10)14 are arginine (R) or lysine (K); Xaai7 is silk Amine acid (S) is either absent; Xaa18* arginine (R), lysine (K) is absent; Xaa19 is phenylalanine (F) or is absent; and Xaa2 is a spermine Acid (R), lysine (K) or is absent. In another aspect, the present invention provides an isolated peptide having a length of from 14 to 50 D-amines; an acid or an L-amino acid, comprising an amino acid sequence of formula V:

XaarXaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaai〇-XaairW-Xaa13 _ -W-Xaai5-Xaai6-Xaai7-LW-Xaa2〇-Xaa2rXaa22 (SEQ ID NO: 595), wherein: 3&amp 1 is tryptophanic acid (W) or is absent; Xaa2 is sulphate 10 or is absent; Xaa3 is lysine (K), face acid (E), arginine (R) , aspartic acid (D) or is absent; Xaa4 is phenylalanine or is absent; Xaas is aspartic acid (D), arginine (R), glutamic acid (E), lysine ( K) is either absent; Xaa0 is serine (S), threonine (T), leucine (L) or is absent; Xaa? is leucine (L) or proline (V) Xaa8 is proline (V), -15 alanine (A) or serine; Xaa9 is glutamic acid (E), lysine (K), scorpion acid (D), sulphonic acid ( T), histidine (H), serine (S) or arginine (R); ^ Xaa10 is cysteine (C), glutamic acid (Ε), leucine (L), silk Barium sulphate or arginine (R); Xaau is isoleucine (I) or valine (V); W is tryptophan; Xaai^aspartic acid (D), arginine (R), Gluten (E) or lysine (K); 20 xaai5 is isoleucine (I) or valine (V); XaaiA aspartic acid (D), arginine (R), bran Amine acid (E), alanine (A), isoleucine (I) or lysine (K); and &&17 is arginine (R), aspartic acid (D), amine Acid (仏), tryptophan 〇^) or glutamic acid (E); L is leucine; (10) is derivatized with serine (S), aspartic acid (D) or threonine (T); And &&21 is glycine (G) or absent; and Xaa22 is serine 9 200837075 acid (s) or is absent; and wherein the amino acid sequence of the formula is not: KFDSLVECIWDWIDRLWS (SEQ ID NO: 96), FDSLVECIWDWIDRLWS (SEQ ID NO: 99), DSLVECIWDWIDRLWS (SEQ ID NO: 100), 5 SLVECIWDWIDRLWS (SEQ ID NO: 101), or LVECIWDWIDRLWS (SEQ ID NO: 102). In several embodiments, the peptide of the invention comprises an amino acid sequence of formula VI:

Xaai-Xaa2_Xaa3_Xaa4-Xaa5-Xaa6-L_V_Xaa9-CIW-Χααΐ3^-Ι-Χαα16-Χαα17-ί^-8-Χαα2ι-Χαα22 (SEQ ID NO: 596), l〇·· Xaai, Xa&2, Xaa2i and Xaa22 In the absence of Xaa3, it is lysine (K), glutamic acid (E), aspartic acid (D), arginine (R) or is absent; Xaa4 is phenylalanine (F) or not Exist; xaa5 is aspartic acid (D), arginine (R), lysine (K) or is absent; further is seric acid (8) or is absent; 15 L is leucine; V is Proline; xaa9 is lysine (K), arginine (R), glutamic acid (E) or aspartic acid (D); C is cysteine; I is isoleucine; W is Tryptophan; Xaa13 is aspartic acid (D), arginine (R) or lysine (K); Xaa10 is arginine (R), aspartic acid (D) or lysine (K); Xaa17 is arginine (R), aspartic acid (D), glutamic acid (E) or lysine (K); and S is serine. In some embodiments, the peptide of the invention comprises an amino acid sequence of formula VII:

Xaa1-Xaa2-Xaa3_Xaa4_Xaa5-Xaa6-L_V_Xaa9-C_I-W_Xaa13-WI-Xaa16-RLWS-Xaa21-Xaa22 (SEQ ID NO: 597), wherein: Xaa!, Xaas, Xaa2AXaa22 are absent; xaa3 is from 10 200837075 amine Acid (K) is either absent; Xaa4 is phenylalanine (F) or is absent; Xaas is arginine (R) or lysine (K) or is absent; xaa6 is serine (s Or in the absence of L, leucine; V is lysine; Xaa9 is lysine (K), face acid (E) or aspartic acid (D); C is cysteine; Isoleucine, w is 5 tryptophan; Xaan is arginine (R) or lysine (κ); xaai6 is aspartic acid (D), arginine (8) or lysine (K); R is arginine; and S is serine. In several embodiments, the peptide of the invention comprises an amino acid sequence of formula VIII:

Xaai-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-LV-Xaa9-CIW-10 Xaai3-WI-Xaai6-Xaai7-LWS-Xaa2i-Xaa22 (SEQ ID NO: 598), where ··· Xaa〗, Xaa!, Xaa2AXaa22 Is absent; xaa3 is arginine (R), lysine (K) or is absent; xaa4 is phenylalanine (F) or is absent; Xaas is arginine (R), lysine (κ) is either absent; xaa6 is serine acid (S) or is absent; L is leucine; V is valine; Xaa9, 15 Xaa13, Xaa^Xaa^ arginine (R) or away Amine acid (κ); C is cysteine; I is isoleucine; W is tryptophan; and S is serine. In several embodiments of the invention, the isolated peptide comprises an amino acid sequence selected from the group consisting of: SWRLDIWDWICESVLDFK (SEQ ID NO: 119), 20 DWLRIIWDWVCSVVSDFK (SEQ ID NO: 123), SWLWEVWDWVLHVLSDFK (SEQ ID NO: 124), TWLRAIWDWVCTALTDFK (SEQ ID NO: 125), SWLRDVWDWVCTVLSDFK (SEQ ID NO: 126), SWLRDIWDWISEVLSDFK (SEQ ID NO: 127), 25 SWLDRIWRWICKVLSRFE (SEQ ID NO: 128)? SWLDDIWDWICEVLSDFE (SEQ ID NO: 129), 11 200837075 SWLRRIWRWICKVLSRFK (SEQ ID NO: 130), SWLKEIWEWICDVLSEFR (SEQ ID NO: 131), SWLKDIWDWICEVLSDFR (SEQ ID NO: 132)? SWLKDIWDWICEVLSDFK (SEQ ID NO: 133), 5 SWLREIWEWICDVLSEFK (SEQ ID NO: 134), SWLREIWEWICEVLSEFK (SEQ ID NO: 135), SWLDRIWRWICKVLSRFE (SEQ ID NO: 136)? SWLDDIWDWICEVLSDFE (SEQ ID NO: 137), SWLRRIWRWICKVLSRFK (SEQ ID NO: 138), 10 SWLRDIWDWIREVLSDFK (SEQ ID NO: 139 ), SWLRDIWDWIEEVLSDFK (SEQ ID NO: 140), SGSWLRDIWDWICEVLSDFK (SEQ ID NO: 141)? GSWLRDIWDWICEVLSDFK (SEQ ID NO: 142)? SWLRDIWDWICEVLSDFKT (SEQ ID NO: 143), 15 SWLRDIWDWICEVLSDFKTW (S EQ ID NO: 144), SWLEKIWKWICRVLSKFD (SEQ ID NO: 165); SWLRKIWKWICEVLSDFK (SEQ ID NO: 166); SWLRDIWDWICKVLSKFK (SEQ ID NO: 167); SWLRRIWRWICEVLSDFK (SEQ ID NO: 168); 20 SWLRDIWDWICRVLSRFK (SEQ ID NO: 169); SWLRRIWDWICRVLSDFK (SEQ ID NO: 170); SWLRKIWDWICKVLSDFK (SEQ ID NO: 171); SWLRRIWDWICEVLSRFK (SEQ ID NO: 172); SWLRKIWDWICEVLSKFK (SEQ ID NO: 173); 25 SWLRDIWRWICRVLSDFK (SEQ ID NO: 174); SWLRDIWKWICKVLSDFK (SEQ ID NO: 175); SWLDRIWDWICRVLSRFK (SEQ ID NO: 176); SWLRDIWDWICKVLSKFK (SEQ ID NO: 177); SWLRDIWRWICKVLSRFK (SEQ ID NO: 179); 30 SWLRDIWKWICKVLSKFK (SEQ ID NO: 180); SWLRKIWKWICEVLSKFK (SEQ ID NO) : 181); SWLRRIWRWICEVLSRFK (SEQ ID NO: 182); SWLRRIWRWICDVLSRFK (SEQ ID NO: 183); SWLRKIWKWICKVLSKFK (SEQ ID NO: 185); 35 SWLRRIWRWICRVLSRFK (SEQ ID NO: 186); 12 200837075 SWLRRIWRWICRVLSRFR (SEQ ID NO: 187 SWLKKIWKWICKVLSKFK (SEQ ID NO: 188); SWRLDIWDWICESVLDF (SEQ ID NO: 189), SWRLDIWDWICESVLD (SEQ ID NO: 190), 5 SWRLDIWDWICESVL (SEQ ID NO: 191), SWRLDIWDWICESV (SEQ ID NO: 192), D WLRIIWDWVCSVVSDF (SEQ ID NO: 193), DWLRII WDWVCSVVSD (SEQ ID NO: 194), DWLRII WDWVCSVVS (SEQ ID NO: 195), 10 DWLRII WDWVCSVV (SEQ ID NO: 196), SWLWEVWDWVLHVLSDF (SEQ ID NO: 197), SWLWEVWDWVLHVLSD (SEQ ID NO: 198), SWLWEVWDWVLHVLS (SEQ ID NO: 199), SWLWEVWDWVLHVL (SEQ ID NO: 200), 15 TWLRAIWDWVCTALTDF (SEQ ID NO: 201), TWLRAIWDWVCTALTD (SEQ ID NO: 202), TWLRAIWDWVCTALT (SEQ ID NO: 203) , TWLRAIWDWVCTAL (SEQ ID NO: 204), SWLRDVWDWVCTVLSDF (SEQ ID NO: 205), 20 SWLRDVWDWVCTVLSD (SEQ ID NO: 206), SWLRDVWDWVCTVLS (SEQ ID NO: 207), SWLRDVWDWVCTVL (SEQ ID NO: 208), SWLRDIWDWISEVLSDF (SEQ ID NO: 209), SWLRDIWDWISEVLSD (SEQ ID NO: 210), 25 SWLRDIWDWISEVLS (SEQ ID NO: 211), SWLRDIWDWISEVL (SEQ ID NO: 212)? SWLDRIWRWICKVLSRF (SEQ ID NO: 213), SWLDRIWRWICKVLSR (SEQ ID NO: 214 ), SWLDRIWRWICKVLS (SEQ ID NO: 215), 30 SWLDRIWRWICKVL (SEQ ID NO: 216), SWLDDIWDWICEVLSDF (SEQ ID NO: 217), SWLDDIWDWICEVLSD (SEQ ID NO: 218), SWLDDIWDWICEVLS (SEQ ID NO: 219), SWLDDIWDWICEVL ( SEQ ID NO: 220), 35 SWLRRIWRWICKVLSRF (SEQ ID NO: 221) 5 13 200837075 SWLRRIWRWICKVLSR (SEQ ID NO: 222), SWLRRIWRWICKVLS (SEQ ID NO: 223)? SWLRRIWRWICKVL (SEQ ID NO: 224), SWLKEIWEWICDVLSEF (SEQ ID NO) : 225)? 5 SWLKEIWEWICDVLSE (SEQ ID NO: 226), SWLKEIWEWICDVLS (SEQ ID NO: 227), SWLKEIWEWICDVL (SEQ ID NO: 228), SWLKDIWDWICEVLSDF (SEQ ID NO: 229), SWLKDIWDWICEVLSD (SEQ ID NO: 230), 10 SWLKDIWDWICEVLS (SEQ ID NO: 231), SWLKDIWDWICEVL (SEQ ID NO: 232), SWLKDIWDWICEVLSDF (SEQ ID NO: 233)? SWLKDIWDWICEVLSD (SEQ ID NO: 234), SWLKDIWDWICEVLS (SEQ ID NO: 235), 15 SWLKDIWDWICEVL (SEQ ID NO: 236), SWLREIWEWICDVLSEF (SEQ ID NO: 237), SWLREIWEWICDVLSE (SEQ ID NO: 238)? SWLREIWEWICDVLS (SEQ ID NO: 239), SWLREIWEWICDVL (SEQ ID NO: 240)? 20 SWLREIWEWICEVLSEF (SEQ ID NO: 591 ), SWLREIWEWICEVLSE (SEQ ID NO: 592), SWLREIWEWICEVLS (SEQ ID NO: 593), SWLREIWEWICEVL (SEQ ID NO: 594), SWLDRIWRWICKVLSRF (SEQ ID NO: 241), 25 SWLDRIWRWICKVLSR (SEQ ID NO: 242), SWLDRIWRWICKVLS ( SEQ ID NO: 243), SWLDRIWRWICKVL (SEQ ID NO: 244), SWLDDIWDWICEVLSDF (SEQ ID NO: 245), SWLDDIWDWICEVLSD (SEQ ID NO: 246), 30 SWLDDIWDWICEVLS (SEQ ID NO: 247), SWLDDIWDWICEVL (SEQ ID NO: 248)? SWLRRIWRWICKVLSRF (SEQ ID NO: 249), SWLRRIWRWICKVLSR (SEQ ID NO: 250), SWLRRIWRWICKVLS (SEQ ID NO: 251)? 35 SWLRRIWRWICKVL (SEQ ID NO: 252), 14 200837075 SWLRDIWDWIREVLSDF (SEQ ID NO: 253), SWLRDIWDWIREVLSD (SEQ) ID NO: 254), SWLRDIWDWIREVLS (SEQ ID NO: 255), SWLRDIWDWIREVL (SEQ ID NO: 256), 5 SWLRDIWDWIEEVLSDF (SEQ ID NO: 257), SWLRDIWDWIEEVLSD (SEQ ID NO: 258), SWLRDIWDWIEEVLS (SEQ ID NO: 259 ), SWLRDIWDWIEEVL (SEQ ID NO: 260)? SGSWLRDIWDWICEVLSDF (SEQ ID NO: 261), 10 SGSWLRDIWDWICEVLSD (SEQ ID NO: 262), SGSWLRDIWDWICEVLS (SEQ ID NO: 263), SGSWLRDIWDWICEVL (SEQ ID NO: 264), GSWLRDIWDWICEVLSDF ( SEQ ID NO: 265), GSWLRDIWDWICEVLSD (SEQ ID NO: 266), 15 GSWLRDIWDWICEVLS (SEQ ID NO: 267), GSWLRDIWDWICEVL (SEQ ID NO: 268), SWLRDIWDWICEVLSDFK (SEQ ID NO: 269), SWLRDIWDWICEVLSDF (SEQ ID NO: 270), SWLRDIWDWICEVLSD (SEQ ID NO: 271), 20 SWLRDIWDWICEVLS (SEQ ID NO: 212), SWLRDIWDWICEVLSDFKT (SEQ ID NO: 273), SWLRDIWDWICEVLSDFK (SEQ ID NO: 274), SWLRDIWDWICEVLSDF (SEQ ID NO: 275), SWLRDIWDWICEVLSD (SEQ ID NO: 276), 25 KFDLVSECIWDWIDLRWS (SEQ ID NO: 277), FDLVSECIWDWIDLRWS (SEQ ID NO: 278), DLVSECIWDWIDLRWS (SEQ ID NO: 279), LVSECIWDWIDLRWS (SEQ ID NO: 280), VSECIWDWIDLRWS (SEQ ID NO) : 281), 30 KFDSVVSCVWDWIIRLWD (SEQ ID NO: 282), FDSVVSCVWDWIIRLWD (SEQ ID NO: 283), DSVVSCVWDWIIRLWD (SEQ ID NO: 284), SVVSCVWDWIIRLWD (SEQ ID NO: 285), VVSCVWDWIIRLWD (SEQ ID NO: 286), 35 KFDSLVHLVWDWVEWLWS (SEQ ID NO: 288), 15 200837075 FDSLVHLVWDWVEWLWS (SEQ ID NO: 289), DSLVHLVWDWVEWLWS (SEQ ID NO: 290) 5 SLVHLVWDWVEWLWS (SEQ ID NO: 291), LVHLVWDWVEWLWS (SEQ ID NO: 292), 5 KFDTLATCVWDWIARLWT (SEQ ID NO: 293)? FDTLATCVWDWIARLWT (SEQ ID NO: 294), DTLATCVWDWIARLWT (SEQ ID NO: 295), TLATCVWDWIARLWT (SEQ ID NO: 296), LATCVWDWIARLWT (SEQ ID NO: 297), 10 KFDSLVTCVWDWVDRLWS (SEQ ID NO: 298)? FDSLVTCVWDWVDRLWS (SEQ ID NO: 299), DSLVTCVWDWVDRLWS (SEQ ID NO: 300), SLVTCVWDWVDRLWS (SEQ ID NO: 301), LVTCVWDWVDRLWS (SEQ ID NO: 302), 15 KFDSLVESI WDWIDRLWS (SEQ ID NO) : 303), FDSLVESIWDWIDRLWS (SEQ ID NO: 304), DSLVESI WDWIDRLWS (SEQ ID NO: 305), SLVESIWDWIDRLWS (SEQ ID NO: 306), LVESIWDWIDRLWS (SEQ ID NO: 307), 20 EFRSLVKCIWRWIRDLWS (SEQ ID NO: 308), FRSLVKCIWRWIRDLWS (SEQ ID NO: 309), RSLVKCIWRWIRDLWS (SEQ ID NO: 310), SLVKCIWRWIRDLWS (SEQ ID NO: 311), LVKCIWRWIRDLWS (SEQ ID NO: 312), 25 EFDSLVECIWDWIDDLWS (SEQ ID NO: 313), FDSLVECIWDWIDDLWS (SEQ ID NO: 314), DSLVECIWDWIDDLWS (SEQ ID NO: 315), SLVECIWDWIDDLWS (SEQ ID NO: 316), LVECIWDWIDDLWS (SEQ ID NO: 317), 30 KFRSLVKCIWRWIRRLWS (SEQ ID NO: 318), FRSLVKCIWRWIRRLWS (SEQ ID NO: 319) RSLVKCIWRWIRRLWS (SEQ ID NO: 320), SLVKCIWRWIRRLWS (SEQ ID NO: 321), LVKCIWRWIRRLWS (SEQ ID NO: 322), 35 RFESLVDCIWEWIEKLWS (SEQ ID NO: 323)? 16 200837075 FESLVDCIWEWIEKLWS (SEQ ID NO: 324), E SLVDCIWEWIEKLWS (SEQ ID NO: 325), SLVDCIWEWIEKLWS (SEQ ID NO: 326), LVDCIWEWIEKLWS (SEQ ID NO: 327), 5 RFDSLVECIWDWIDKLWS (SEQ ID NO: 328), FDSLVECIWDWIDKLWS (SEQ ID NO: 329), DSLVECIWDWIDKLWS (SEQ ID NO: 330), SLVECIWDWIDKLWS (SEQ ID NO: 331), LVECIWDWIDKLWS (SEQ ID NO: 332), 10 KFDSLVECIWDWIDKLWS (SEQ ID NO: 333), FDSLVECIWDWIDKLWS (SEQ ID NO: 334), DSLVECIWDWIDKLWS (SEQ ID NO: 335) , SLVECIWDWIDKLWS (SEQ ID NO: 336), LVECIWDWIDKLWS (SEQ ID NO: 337), 15 KFESLVDCIWEWIERLWS (SEQ ID NO: 338), FESLVDCIWEWIERLWS (SEQ ID NO: 339), ESLVDCIWEWIERLWS (SEQ ID NO: 340), SLVDCIWEWIERLWS (SEQ ID NO: 341), LVDCIWEWIERLWS (SEQ ID NO: 342), 20 KFESLVECIWEWIERLWS (SEQ ID NO: 343), FESLVECIWEWIERLWS (SEQ ID NO: 344), ESLVECIWEWIERLWS (SEQ ID NO: 345), SLVECIWEWIERLWS (SEQ ID NO: 346 ), LVECIWEWIERLWS (SEQ ID NO: 347), 25 EFRSLVKCIWRWIRDLWS (SEQ ID NO: 348), FRSLVKCIWRWIRDLWS (SEQ ID NO: 349), RSLVKCIWRWIRDLWS (SEQ ID NO: 350)? SLVKCIWRWIRDLWS (SEQ ID NO: 351), LVKCIWRWIRDLWS ( SEQ ID NO: 35 2), 30 EFDSLVECIWDWIDDLWS (SEQ ID NO: 353), FDSLVECIWDWIDDLWS (SEQ ID NO: 354), DSLVECIWDWIDDLWS (SEQ ID NO: 355), SLVECIWDWIDDLWS (SEQ ID NO: 356), LVECIWDWIDDLWS (SEQ ID NO: 357), 35 KFRSLVKCIWRWIRRLWS (SEQ ID NO: 358), 17 200837075 FRSLVKCIWRWIRRLWS (SEQ ID NO: 359), RSLVKCIWRWIRRLWS (SEQ ID NO: 360) 3 SLVKCIWRWIRRLWS (SEQ ID NO: 361), LVKCIWRWIRRLWS (SEQ ID NO: 362), 5 KFDSLVERIWDWIDRLWS ( SEQ ID NO: 363), FDSLVERIWDWIDRLWS (SEQ ID NO: 364), DSLVERIWDWIDRLWS (SEQ ID NO: 365), SLVERIWDWIDRLWS (SEQ ID NO: 366), LVERIWDWIDRLWS (SEQ ID NO: 367), 10 KFDSLVEEIWDWIDRLWS (SEQ ID NO: 368), FDSLVEEIWDWIDRLWS (SEQ ID NO: 369), DSLVEEIWDWIDRLWS (SEQ ID NO: 370), SLVEEIWDWIDRLWS (SEQ ID NO: 371), LVEE1WDWIDRLWS (SEQ ID NO: 372), 15 KFDSLVECIWDWIDRLWSGS (SEQ ID NO: 373), FDSLVECIWDWIDRLWSGS (SEQ ID NO: 374), DSLVECIWDWIDRLWSGS (SEQ ID NO: 375), SLVECIWDWIDRLWSGS (SEQ ID NO: 376)? LVECIWDWIDRLWSGS (SEQ ID NO: 377), 20 KFDSLVECIWDWIDRLWSG (SEQ ID NO: 378), FDSLV ECIWDWIDRLWSG (SEQ ID NO: 379), DSLVECIWDWIDRLWSG (SEQ ID NO: 380), SLVECIWDWIDRLWSG (SEQ ID NO: 381), LVECIWDWIDRLWSG (SEQ ID NO: 382), 25 TKFDSLVECIWDWIDRLWS (SEQ ID NO: 383), KFDSLVECIWDWIDRLWS (SEQ ID NO: 384), FDSLVECIWDWIDRLWS (SEQ ID NO: 385), DSLVECIWDWIDRLWS (SEQ ID NO: 386), SLVECIWDWIDRLWS (SEQ ID NO: 387), 30 WTKFDSLVECIWDWIDRLWS (SEQ ID NO: 388), TKFDSLVECIWDWIDRLWS (SEQ ID NO: 389) , KFDSLVECIWDWIDRLWS (SEQ ID NO: 390), FDSLVECIWDWIDRLWS (SEQ ID NO: 391), DSLVECIWDWIDRLWS (SEQ ID NO: 392)? SWLEKIWKWICRVLSKF (SEQ ID NO: 393); 18 35 200837075 5 10 15 20 25 30 SWLEKIWKWICRVLSK (SEQ ID NO: 394); SWLEKIWKWICRVLS (SEQ ID NO: 395); SWLEKIWKWICRVL (SEQ ID NO: 396); SWLRKIWKWICEVLSDF (SEQ ID NO: 397); SWLRKIWKWICEVLSD (SEQ ID NO: 398); SWLRKIWKWICEVLS (SEQ ID NO: 399); SWLRKIWDWICE VL (SEQ ID NO: 401); SWLRDIWDWICKVLSK (SEQ ID NO: 402); SWLRDIWDW1CKVLS (SEQ ID NO: 403); SWLRDIWDWICKVL (SEQ ID NO: 404); SWLRRIWRWICEVLSDF (SEQ) ID NO: 405); SWLRR1WRWICEVLSD (SEQ ID NO: 406); SWLRR1WRWICEVLS (SEQ ID NO: 407); SWLRRIWRWICEVL (SEQ ID NO: 408); SWLRDIWDWICRVLSRF (SEQ ID NO: 409); SWLRDIWDWICRVLSR (SEQ ID NO: 410) SWLRDIWDWICRVLS (SEQ ID NO: 411); SWLRDIWDWICRVL (SEQ ID NO: 412); SWLRRIWDWICRVLSDF (SEQ ID NO: 413); SWLRRIWDWICRVLSD (SEQ ID NO: 414); SWLRRIWDWICRVLS (SEQ ID NO: 415); SWLRRIWDWICRVL (SEQ ID NO: 416); SWLRKIWDWICKVLSDF (SEQ ID NO: 417); SWLRKIWDWICKVLSD (SEQ ID NO: 418); SWLRKIWDWICKVLS (SEQ ID NO: 419); SWLRKIWDWICKVL (SEQ ID NO: 420); SWLRRIWDWICEVLSRF (SEQ ID NO: 421); SWLRRIWDWICEVLSR (SEQ ID NO: 422); SWLRRIWDWICEVLS (SEQ ID NO: 423); SWLRRIWDWICEVL (SEQ ID NO: 424); SWLRKIWDWICEVLSKF (SEQ ID NO: 425); SWLRKIWDW1CEVLSK (SEQ ID NO: 426); SWLRKIWDWICEVLS (SEQ ID NO) : 427); SWLRKIWDWICEVL (SEQ ID NO: 428); 19 35 200837075 SWLRDIWRWICRVLSDF (SEQ ID NO: 429); SWLRDIWRWICRVLSD (SEQ ID NO: 430); SWLRDIWRWICRVLS (SEQ ID NO: 431); SWLRDIWRWICRVL (SEQ ID NO: 432 ); 5 SWLRDIWKWICKVLSDF (SEQ ID NO: 433); SWL RDIWKWICKVLSD (SEQ ID NO: 434); SWLRDIWKWICKVLS (SEQ ID NO: 435); SWLRDIWKWICKVL (SEQ ID NO: 436); SWLDR1WDWICRVLSRF (SEQ ID NO: 437); 10 SWLDRIWDWICRVLSR (SEQ ID NO: 438); SWLDRIWDWICRVLS (SEQ ID NO: 439); SWLDRIWDWICRVL (SEQ ID NO: 440); SWLRDIWDWICKVLSKF (SEQ ID NO: 441); SWLRDIWDWICKVLSK (SEQ ID NO: 442); 15 SWLRDIWDWICKVLS (SEQ ID NO: 443); and SWLRDIWDWICKVL (SEQ ID NO: 444 ). SWLRDIWRWICKVLSRF (SEQ ID NO: 445); SWLRDIWRWICKVLSR (SEQ ID NO: 446); SWLRDIWRWICKVLS (SEQ ID NO: 447); 20 SWLRDIWRWICKVL (SEQ ID NO: 448); SWLRDIWKWICKVLSKF (SEQ ID NO: 449); SWLRDIWKW1CKVLSK (SEQ ID NO: 450); SWLRDIWKWICKVLS (SEQ ID NO: 451); SWLRDIWKWICKVL (SEQ ID NO: 452); 25 SWLRKIWKWICEVLSKF (SEQ ID NO: 453); SWLRKIWKWICEVLSK (SEQ ID NO: 454); SWLRKIWKWICEVLS (SEQ ID NO: 455) SWLRKIWKWICEVL (SEQ ID NO: 456); SWLRRIWRWICEVLSRF (SEQ ID NO: 457); 30 SWLRRIWRWICEVLSR (SEQ ID NO: 458); SWLRRIWRWICEVLS (SEQ ID NO: 459); SWLRRIWRWICEVL (SEQ ID NO: 460); SWLRRIWRWICDVLSRF (SEQ ID NO: 461); SWLRRIWRWICDVLSR (SEQ ID NO: 462); 35 SWLRRIWRWICDVLS (SEQ ID NO: 463), 20 200837075 SWLRRIWRWICDVL (SEQ ID NO: 464), SWLRKIWKWICKVLSKF (SEQ ID NO: 465); SWLRKIWKWICKVLSK (SEQ ID NO) : 466); SWLRKIWKWICKVLS (SEQ ID NO: 467); 5 SWLRKIWKWICKVL (SEQ ID NO: 468); SWLRRIWRWICRVLSRF (SEQ ID NO: 469); SWLRRIWRWICRVLSR (SEQ ID NO: 470); SWLRRIWRWICRVLS (SEQ ID NO: 471); SWLRRIWRWICRVL (SEQ ID NO: 472); 10 SWLRRIWRWICRVLSRF (SEQ ID NO: 473); SWLRRIWRWICRVLSR (SEQ ID NO: 474); SWLRRIWRWICRVLS (SEQ ID NO: 475); SWLRRIWRWICRVL (SEQ ID NO: 476); SWLKKIWKWICKVLSKF (SEQ ID NO: 477); 15 SWLKKIWKWICKVLSK (SEQ ID NO: 478); SWLKKIWKWICKVLS (SEQ ID NO: 479); SWLKKIWKWICKVL (SEQ ID NO: 480); DFKSLVRCIWKWIKELWS (SEQ ID NO: 481); FKSLVRCIWKWIKELWS (SEQ ID NO: 482); 20 KSLVRCIWKWIKELWS (SEQ ID NO: 483) SLVRCIWKWIKELWS (SEQ ID NO: 484); LVRCIWKWIKELWS (SEQ ID NO: 485); KFDSLVECIWKWIKRLWS (SEQ ID NO: 486); FDSLVECIWKWIKRLWS (SEQ ID NO: 487); 25 DSLVECIWKWIKRLWS (SEQ ID NO: 488); SLVECIWKWIKRLWS (SEQ ID NO: 489); LVECIWKWIKRLWS (SEQ ID NO: 490); KFKSLVKCIWDWIDRLWS (SEQ ID NO: 491); FKSLVKCIWDWIDRLWS (SEQ ID NO: 492); 30 KSLVKCIWDWIDRLWS (SEQ ID NO: 493); SLVKCIWDWIDRLWS (SEQ ID NO: 494 LVKCIWDWIDRLWS (SEQ ID NO: 495); KFDSLVECIWRWIRRLWS (SEQ ID NO: 496); FDSLVECIWRWIRRLWS (SEQ ID NO: 497); DSLVECIWRWIRRLWS (SEQ ID NO: 498); 21 35 200837075 SLVECIWRWIRRLWS (SEQ ID NO: 499); LVECIWRWIRRLWS (SEQ ID NO: 50 0); KFRSLVRCIWDWIDRLWS (SEQ ID NO: 501); FRSLVRCIWDWIDRLWS (SEQ ID NO: 502); 5 RSLVRCIWDWIDRLWS (SEQ ID NO: 503); SLVRCIWDWIDRLWS (SEQ ID NO: 504); LVRCIWDWIDRLWS (SEQ ID NO: 505); KFDSLVRCIWDWIRRLWS (SEQ ID NO: 506); FDSLVRCIWDWIRRLWS (SEQ ID NO: 507); 10 DSLVRCIWDWIRRLWS (SEQ ID NO: 508); SLVRCIWDWIRRLWS (SEQ ID NO: 509); LVRCIWDWIRRLWS (SEQ ID NO: 510); KFDSLVKCIWDWIKRLWS (SEQ ID NO) : 511); FDSLVKCIWDWIKRLWS (SEQ ID NO: 512); 15 DSLVKCIWDWIKRLWS (SEQ ID NO: 513); SLVKCIWDWIKRLWS (SEQ ID NO: 514); LVKCIWDWIKRLWS (SEQ ID NO: 515); KFRSLVECI WDWIRRLWS (SEQ ID NO: 516); FRSLVECIWDWIRRLWS (SEQ ID NO: 517); 20 RSLVECIWDWIRRLWS (SEQ ID NO: 518); SLVECIWDWIRRLWS (SEQ ID NO: 519); LVECIWDWIRRLWS (SEQ ID NO: 520); KFKSLVECIWDWIKRLWS (SEQ ID NO: 521); FKSLVECIWDWIKRLWS (SEQ ID NO: 522); 25 KSLVECIWDWIKRLWS (SEQ ID NO: 523); SLVECIWDWIKRLWS (SEQ ID NO: 524); LVECIWDWIKRLWS (SEQ ID NO: 525); KFDSLVRCIWRWIDRLWS (SEQ ID NO: 526); FDSLVRCIWRWIDRLWS (SEQ ID NO: 527) ; 30 DSLVRCIWRWIDRLWS (SEQ ID NO: 52 8); SLVRCIWRWIDRLWS (SEQ ID NO: 529); LVRCIWRWIDRLWS (SEQ ID NO: 530); KFDSLVKCIWKWIDRLWS (SEQ ID NO: 531); FDSLVKCIWKWIDRLWS (SEQ ID NO: 532); DSLVKCIWKWIDRLWS (SEQ ID NO: 533); 200837075 SLVKCIWKWIDRLWS (SEQ ID NO: 534); 5 10 LVKCIWKWIDRLWS (SEQ ID NO: 535); KFRSLVRCIWDWIRDLWS (SEQ ID NO: 536); FRSLVRCIWDWIRDLWS (SEQ ID NO: 537); RSLVRCIWDWIRDLWS (SEQ ID NO: 538); SLVRCIWDWIRDLWS ( SEQ ID NO: 539); LVRCIWDWIRDLWS (SEQ ID NO: 540); KFKSLVKCIWDWIDRLWS (SEQ ID NO: 541); FKSLVKCIWDWIDRLWS (SEQ ID NO: 542); KSLVKCIWDWIDRLWS (SEQ ID NO: 543); SLVKCIWDWIDRLWS (SEQ ID NO: 544 LVKCIWDWIDRLWS (SEQ ID NO: 545); KFRSLVKCIWRWIDRLWS (SEQ ID NO: 546); 15 20 25 FRSLVKC1WRWIDRLWS (SEQ ID NO: 547); RSLVKCIWRWIDRLWS (SEQ ID NO: 548); SLVKCIWRWIDRLWS (SEQ ID NO: 549); LVKCIWRWIDRLWS (SEQ ID NO: 550); KFKSLVKCIWKWIDRLWS (SEQ ID NO: 551); FKSLVKCIWKWIDRLWS (SEQ ID NO: 552); KSLVKCIWKWIDRLWS (SEQ ID NO: 553); SLVKCIWKWIDRLWS (SEQ ID NO: 554); LVKCIWKWIDRLWS (SEQ ID NO) : 555); KFKSLVECIWKWIKRLWS (SEQ ID NO: 556); FKSLVECIWKWIKRLWS (SEQ ID NO: 557); KSLVECIWKWIKRLWS (SEQ ID NO: 558); SLVECIWKWIKRLWS (SEQ ID NO: 559); LVECIWKWIKRLWS (SEQ ID NO: 560); KFRSLVECIWRWIRRLWS (SEQ ID NO: 561) FRSLVECIWRWIRRLWS (SEQ ID NO: 562); 30 RSLVECIWRWIRRLWS (SEQ ID NO: 563); SLVECIWRWIRRLWS (SEQ ID NO: 564); LVECIWRWIRRLWS (SEQ ID NO: 565); KFRSLVDCIWRWIRRLWS (SEQ ID NO: 566). FRSLVDCIWRWIRRLWS (SEQ ID NO: 567). RSLVDCIWRWIRRLWS (SEQ ID NO: 568). 23 35 200837075 SLVDCIWRWIRRLWS (SEQ ID NO: 569). LVDCIWRWIRRLWS (SEQ ID NO: 570). KFKSLVKCIWKWIKRLWS (SEQ ID NO: 571); FKSLVKCIWKWIKRLWS (SEQ ID NO: 572); 5 KSLVKCIWKWIKRLWS (SEQ ID NO: 573); SLVKCIWKWIKRLWS (SEQ ID NO: 574); LVKCIWKWIKRLWS (SEQ ID NO: 575); KFRSLVRCIWRWIRRLWS (SEQ ID NO: 576); FRSLVRCIWRWIRRLWS (SEQ ID NO: 577); 10 RSLVRCIWRWIRRLWS (SEQ ID NO: 578); SLVRCIWRWIRRLWS (SEQ ID NO: 579); LVRCIWRWIRRLWS (SEQ ID NO: 580); RFRSLVRCIWRWIRRLWS (SEQ ID NO: 581) ; FRSLVRCIWRWIRRLWS (SEQ ID NO: 582); 15 RSLVRCIWRWIRRLWS (SEQ ID NO: 583); SLVRCIWRWIRRLWS (SEQ ID NO: 584); LVRCIWRWIRRLWS (SEQ ID NO: 585); KFKSLVKCIWKWIKKLWS (SEQ ID NO: 586), FKSLVKCIWKWIKKLWS (SEQ ID NO: 587), 20 KSLVKCIWKWIKKLWS (SEQ ID NO: 588), SLVKCIWKWIKKLWS (SEQ ID NO: 589), LVKCIWKWIKKLWS (SEQ ID NO: 590). In another embodiment of the invention, the isolated peptide of the invention is derived from any of the aforementioned amino acid sequences, in particular, SEQ ID NO: 25 119, 123 to 144, 165 to 177; 179 to 183; 185 to 590; and 591 to 594. In several embodiments, the peptide of the invention has virucidal activity. In several embodiments, the peptide of the present invention is 18 to 40 D-amino acids or L. amino acids, 18 to 30 D-amino acids or L-amino acids, or long. 18 30 to 22 D-amino acids or L-amino acids. In several embodiments, the peptide of the present invention comprises a D-amino acid. In other embodiments, the peptide of the invention comprises 24 200837075 L-amino acid. In several embodiments, the peptide comprises a dimethylaminonaphthalene moiety. In several embodiments, the peptide has an EC5G of about 3 μM or less; about 2 μM or less, about ΙμΜ or less, about 5 〇〇ηΜ or less; about 400 Μ 或 or less; or about 300 Μ. In several embodiments, the peptides are active against HIV, hepatitis 5 c virus or a flavivirus such as West Nile virus or dengue virus. In another aspect, the invention provides a pharmaceutical composition comprising any of the peptides of the invention as described herein. In several embodiments, the pharmaceutical composition is a microbicide or a vaginal cream. In the U.S. embodiment, the pharmaceutical composition comprises an antiviral agent. In several embodiments, the sputum agent is a protease inhibitor, a polymerase inhibitor, a ligase inhibitor, an invasion inhibitor, an assembly/secretion inhibitor, a translation inhibitor, or an immunostimulatory agent. In several embodiments, the antiviral agent is interferon, pegylated interferon, ribavirin 'amantadine, rimanta (iine), piconari (pleconaril), acyclovir, zidovudine, lamivudine, Indenavir (Merck), Tarapuvi ( Telaprivir) (Vertex), Tenofivir (Gilead), R1626 (R0Che), GS-9137 (Gee 20), Fuzeon ) (Roche, Trimeris, Celgosivir (Migenix), VGX-410c (VGX Pharmaceuticals), 0 IMO-2125 (Indira Pharmaceuticals) Idem pharmaceuticals)) or a combination thereof. In another aspect, the present invention provides a manufactured article comprising a 25 200837075 container for collecting body fluids and any of the advantages of the invention described herein, in several embodiments, The container is a collection bag, tube, capillary or, main shot. 5 10 15 20 In several embodiments, the container Vacuuming. In several embodiments, the container further comprises a bio-stabilizer. In several embodiments, the peptide is attached to the container or adsorbed to the container, so that the material is placed therein; The peptide is retained in the container. When the peptide is attached to the crying = adsorbed on the container, the peptide can still inhibit the viral infection. ^ In the other aspect, the present invention provides a composition Included is a sample obtained from a mammalian body and any of the peptides of the invention as described herein. In several embodiments, the composition also includes a biological stabilizer. In several embodiments, the sample is a blood product. In some embodiments, the blood product is plasma, platelets, white blood cells, or stem cells. In the composition or article of the present invention, the biological stabilizer is an anticoagulant, a preservative, a protease inhibitor, or a combination thereof In some embodiments, the anticoagulant is citrate, ethyldiaminetetraacetic acid (EDTA), heparin, oxalate, fluoride, or a combination thereof. In several embodiments, Preservative Boric acid, sodium formate and sodium borate. In several embodiments, the protease inhibitor is di-peptidyl protease (7). In several embodiments, the peptide and/or stabilizer are lyophilized. The present invention provides (1) a method for deactivating a virus, relating to contacting the virus with any of the peptides of the present invention described herein; (2) a method for preventing or treating a mammalian cell infected with a virus, involving Mammalian cells are contacted with any of the peptides of the invention described herein or (3) - a method of preventing or treating a viral infection in a mammalian body, 26 200837075 relates to any one of an effective amount administered to the mammal A peptide of the invention as described herein. In several embodiments of the methods of the invention, the virus is a flavivirus, a hepatic virus or a human immunodeficiency virus. In several embodiments, the virus is c5 hepatitis virus, West Nile virus or dengue virus. In several embodiments, the mammalian cell is a human cell. In several embodiments, the mammal is a human. In several embodiments, the peptide is administered topically or systemically. In another aspect, the invention provides U) a method of deactivating a human immune 10 deficiency virus; (2) a method of preventing or treating a mammalian cell infected with a human immunodeficiency virus; and a method of preventing or treating a mammalian human Immunization lacks a method of viral infection. Such methods involve contacting the mother or mammalian cells, or administering to the mammal 4 to 5 D- or L-amino acids, wherein the peptide comprises a helix The structure, 15 and wherein the polar amino acid is on the same side of the alpha-helical structure' and the non-polar amino acid is on the other side of the helical structure. For example, all of the polar amino acids are located on the same side of the helical structure, and all of the non-polar amino acids are located on the other side of the helical structure, resulting in an intact amphiphilic property. As is well known in the art, the intact amphiphilic nature can be determined using a spiral wheel. In several embodiments, the method also involves contacting the virus or cell with a pharmaceutical composition of the invention. In several embodiments, the mammalian cell is a human cell. In several embodiments, the peptide is also effective to treat, inhibit or prevent infection by a combination of hepatitis C virus, West Nile virus, dengue virus or its 2008 200837075. In several embodiments, a peptide useful in the methods of the invention, such as selected from the group consisting of alanine, valine, leucine, methionine, isoleucine, phenylalanine, and tryptamine A group of non-polar amino acids consisting of acids. 5 In several embodiments, the polar amino acid is selected from the group consisting of arginine, aspartame, aspartic acid, cysteine, glutamic acid, glutamine, histidine, homocysteine A group consisting of acid, lysine, hydroxy lysine, ornithine, serine and threonine. In several embodiments, the cysteine residue of the peptide is at the N-terminal position of the peptide of the peptide. In several embodiments, the cysteamine 10 acid is located at four positions on the N-terminus of the peptide of the peptide. In several embodiments, the cysteine acid is located at a position relative to the N-terminus of the peptide. In several embodiments, the amino acid 16 and the amino acid 18 are charged with respect to the N-terminus of the peptide. Specifically, the amino acid 16 and the amino acid 18 are positively and negatively charged, positively and positively, respectively. Electrical, or negative and positive. 15 々 A number of real-cut towels, which can be used in the method of the present invention - the peptide is 14 to 40 D_ or L-amino acids, 14 to 3 long, 1) or 1 amino acid. , 14 to 25 D- or L-amino acids, or 14 to 18 〇 or _ amino acids. In several embodiments, the peptide is 14 amino acids long; the amino acids are arginine, cysteine, glutamic acid, serine, proline, and two days of 20-butic acid. , two leucine, two isoleucine, and three tryptophan residues; and the arginine, cysteine, amylin, serine, and aspartate residues Located on the same side of the alpha-helical structure. In other embodiments, the peptide is 15 amino acids long; the amino acids are arginine, cysteine, glutamic acid, two serines, proline, and two aspartic acid , two leucine, 28 200837075 two isoleucine, and three tryptophan residues; and the arginine, cysteine, glutamic acid, serine, and aspartate The base is located on the same side of the alpha-helical structure. In other embodiments, the peptide is 16 amino acids in length; the amino acids are arginine, cysteine, glutamic acid, two serines, 5 valine, three winters Acid, two leucine, two isoleucine, and three tryptophan residues; and the arginine, cysteine, glutamic acid, serine, and aspartate residues It is located on the same side of the spiral structure. In several embodiments, the peptide is 17 amino acids in length; the amino acids are arginine, cysteine, glutamic acid, two serines, valine, three days 10 Acid, two leucine, two isoleucine, three tryptophan and phenylpropylamine oxime, and far-inspired arginine, semi-deaminic acid, face acid, serine, And the aspartic acid residue is located on the same side of the α-helical structure. In several embodiments, the peptide is 18 amino acids long; the amino acids are arginine, cysteine, glutamic acid, two serines, valine, and three aspartic acid , two 15 leucine, two isoleucine, three tryptophan, phenylalanine and lysine residues; and the arginine, cysteine, glutamic acid, silkamine The acid and aspartic acid residues are on the same side of the alpha-helical structure. In several embodiments, the peptide used in the methods of the invention consists of a D-amino acid. In other embodiments, the peptide consists of L-amino acid. In a further embodiment 2, a dimethylaminonaphthalenesulfonyl moiety is further included. In several embodiments, a peptide useful in the methods of the invention comprises the following amino acid sequence: QIVGGVYLLPRRGPRLGV (SEQ ID NO: 4), QPGYPWPLYGNEGCGWAG (SEQ ID NO: 5), 29 200837075 LYGNEGCGWAGWLLSPRG (SEQ ID NO: 6) GWAGWLLSPRGSRPSWGP (SEQ ID NO: 7)? IFLLALLSCLTVPASAYQ (SEQ ID NO: 8), DAILHTPGCVPCVREGNA (SEQ ID NO: 9), 5 LPTTQLRRHIDLLVGSAT (SEQ ID NO: 10), RHIDLLVGSATLCSALYV (SEQ ID NO: 11)? GSATLCSALYVGDLCGSV (SEQ ID NO: 12), ALYVGDLCGSVFLVGQLF (SEQ ID NO: 13), IMDMIAGAHWGVLAGIAY (SEQ ID NO: 14), 10 HINSTALNCNESLNTGWL (SEQ ID NO: 15), NCNESLNTGWLAGLFYQH (SEQ ID NO: 16), LASCRRLTDFAQ GWGPIS (SEQ ID NO: 17 ), TDFAQGWGPISYANGSGL (SEQ ID NO: 18), GPISYANGSGLDERPYCW (SEQ ID NO: 19), 15 GSGLDERPYCWHYPPRPC (SEQ ID NO: 20), WMNSTGFTKVCGAPPCVI (SEQ ID NO: 21)? PCVIGGVGNNTLLCPTDC (SEQ ID NO: 22), MYVGGVEHRLEAACNWTR ( SEQ ID NO: 23), YLYGVGSSIASWAIKWEY (SEQ ID NO: 24), 20 SIASWAIKWEYVVLLFLL (SEQ ID NO: 25), KWEYVVLLFLLLADARVC (SEQ ID NO: 26), WMMLLISQAEAALENLVI (SEQ I D NO: 27), GAVYAFYGMWPLLLLLLA (SEQ ID NO: 28), GMWPLLLLLLALPQRAYA (SEQ ID NO: 29), 25 TLVFDITKLLLAIFGPLW (SEQ ID NO: 30), VSTATQTFLATCIN (SEQ ID NO: 31), ATQTFLATCINGVCWTVY (SEQ ID NO: 32 ), DSSVLCECYDAGCAWYEL (SEQ ID NO: 33), AYMNTPGLPVCQDHLEFW (SEQ ID NO: 34), 30 LEFWEGVFTGLTHIDAHF (SEQ ID NO: 35), HPITKYIMTCMSADLEVV (SEQ ID NO: 36), VTSTWVLVGGVLAAL (SEQ ID NO: 37), WVLVGGVLAALAAYCLST ( SEQ ID NO: 38), LAALAAYCLSTGCVV (SEQ ID NO: 39), 35 EVFWAKHMWNFISGIQYL (SEQ ID NO: 40)? 30 200837075 MWNFISGIQYLAGLSTLP (SEQ ID NO: 41), PAILSPGALVVGVVCAAI (SEQ ID NO: 42)? SWLRDIWDWICEVLSDFK (SEQ ID NO: 43), DWICEVLSDFKTWLKAKL (SEQ ID NO: 44), 5 YVSGMTTDNLKCPCQIPS (SEQ ID NO: 45), SSGADTEDVVCCSMS (SEQ ID NO: 46), DTEDVVCCSMSYSW (SEQ ID NO: 47), SSGADTEDVVCCSMSYSW (SEQ ID NO: 48) , DVVCCSMSYSWTGAL (SEQ ID NO: 49)? 10 TVTESDIRTEEAIYQCCD (SEQ ID NO: 50), GNTLTCYIKARAACRAAG (SEQ ID NO: 51), RAAGLQDCTMLVCGDDLV (SEQ ID NO: 52)? CTMLVCGDDLVVICESAG (SEQ ID NO: 53), DDLVVI CESAGVQEDAAS (SEQ ID NO: 54), 15 LELITSCSSNVSVAHDGA (SEQ ID NO: 55), HTPVNSWLGNIIMFAPTL (SEQ ID NO: 56), APTLWARMILMTHFFSVL (SEQ ID NO: 57), DQLEQALNCEIYGACYSI (SEQ ID NO: 58), GVPPLRAWRHRARSVRAR (SEQ ID NO: 59), 20 WRHRARSVRARLLSRGGR (SEQ ID NO: 60), GWFTAGYSGGDIYHSVSH (SEQ ID NO: 61), LYGNEGLGWAGWLLSPRG (SEQ ID NO: 62)? IFLLALLSCITVPVSAAQ (SEQ ID NO: 63), IFLLALLSCLTIPASAYE (SEQ ID NO: 64) , 25 MSATFCSALYVGDLCGGV (SEQ ID NO: 65), GAAALCSAMYVGDLCGSV (SEQ ID NO: 66), ALYVGDLCGGVMLAAQVF (SEQ ID NO: 67), AMYVGDLCGSVFLVAQLF (SEQ ID NO: 68), IIDIVSGAHWGVMFGLAY (SEQ ID NO: 69), 30 VVDMVAGAHWGVLAGLAY ( SEQ ID NO: 70), VDVQYMYGLSPAITKYVV (SEQ ID NO: 71), YLYGIGSAVVSFAIKWEY (SEQ ID NO: 72)? WMLILLGQAEAALEKLVV (SEQ ID NO: 73)? WMMLLIAQAEAALENLVV (SEQ ID NO: 74), 35 GVVFDITKWLLALLGPAY (SEQ ID NO: 75)9 31 200837075 ELIFTITK1LLAILGPLM (SEQ ID NO:76)? VSQSFLGTTISGVLWTVY (SEQ ID NO:77)5 ATQSFLATCVNGVCWTVY (SEQ ID NO:78), SWLRDVWDWVCTILTDFK (SEQ ID NO:79), 5 SWLRDVWDWICTVLTDFK (SEQ ID NO: 80) 5 DWVCTILTDFKNWLTSKL (SEQ ID NO: 81)? DWICTVLTDFKTWLQSKL (SEQ ID NO: 82), ASEDVYCCSMSYTWT (SEQ ID NO: 83), EDDTTVCCSMSYSW (SEQ ID NO: 84), 10 CTMLVCGDDLVVICESAG (SEQ ID NO: 85) , PTMLVCG DDLVVISESQG (SEQ ID NO: 86), SWLRPIWPWICEVLSDFK (SEQ ID NO: 91), SWLRDIWDWICEVL (SEQ ID NO: 92), SWLRDIWDWICEVLS (SEQ ID NO: 93), 15 SWLRDIWDWICEVLSD (SEQ ID NO: 94), SWLRDIWDWICEVLSDF ( SEQ ID NO: 95), KFDSLVECIWDWIDRLWS (SEQ ID NO: 96), SIWRDWVDLICEFLSDWK (SEQ ID NO: 97), KWLCRIWSWISDVLDDFE (SEQ ID NO: 98), 20 FDSLVECIWDWIDRLWS (SEQ ID NO: 99), DSLVECIWDWIDRLWS (SEQ ID NO: 100) SLVECIWDWIDRLWS (SEQ ID NO: 101), or LVECIWDWIDRLWS (SEQ ID NO: 102) In several embodiments, the peptides useful in the methods of the invention are SEQ 25 ID NO.. 4-86 and 91-102 Any of a group of amino acid sequences. In the examples, the peptide useful in the method of the present invention has an EC5 of about 3 μM or less, about 2 μM or less, about ΙμΜ or less, about 500 Å or less, about 400 Å or less, about 300 Å. In another aspect, the present invention provides an isolated peptide having from 14 to 50 D- or L-amine 30-acids having an amphipathic a-helical structure and having an anti-HIV and/or flavivirus Antiviral activity of the virus. 32 200837075 In one embodiment, the peptide has a sequence comprising any one of Formulas IX-XIII:

Xaai-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8- IX

Xaa9-Xaai〇rXaairXaai2-Xaai3-Xaai4 (SEQ EDNO: 112)

Xaa! -Xaa2_Xaa3 -Xa34-Xaa5-Xa〇5-Xaa7,Xaa8-Xaa9- X

Xaaio-Xaan-Xaan-XaaB-XaaH-Xaais (SEQIDNO: 113)

v Xaai_Xaa2_XHE3-XaB4-Xaa5-Xa35-Xaa7-Xaa8-Xaa9-XB34〇- XI 10 X〇Hn-Xaai2-Xaai3-Xaai4· 46 (SEQ ID NO: 114)

(10)(10)7-Xd3^-Xaap-Xadi 〇-Xaai i - XII

Xaai2-Xaai3-Xaai4-Xaai5-Xaai6-Xaai7 (SEQIDNO: 115) 15

Xaaj -Xaa2-Xaa3-XaB4-Xaa5-Xa3^-Xaa7 - Xaa8_Xaa9-Xaai 〇_Xaai i -

Xaa!2_X(10)ΐ3_Χ^^ΐ4·Χ^ΐ5-Χ^ΐ6-Χ^ΐ7-Xaai8 (SEQ ID NO: 116) wherein: Xaai, Xaa4, Xaa5, Xaa8, Xaan, Xaa12, Xaa15, Xaa16AXaa18 are each a polar amino acid; And xaa2, xaa3, xaa6, Xaa7, Xaa9, Xaa1(), Xaa13, Xaa14, and 17 are each a non-polar 20 amino acid. In another embodiment, the invention provides a fusion peptide formed by attaching a 14 amino acid peptide (N-terminal peptide) to the n-terminus of any of the formulae IX to XIII. The 14 amino acid N-terminal peptide has the structural formula:

Rx-Ry-Ry-Rx-Ry-Ry-Rx-Rx-Ry-Ry-Rx.Rx.Ry.Rx (SEQ ID 25 NO: 117), wherein each Rx is a polar amino acid, and each _ separately It is a non-polar amino acid. In another embodiment, the present invention provides a fusion formed by the attachment of a 12 amino acid peptide (C-terminal peptide) to the c-terminus of the acesulfonin 33 200837075 peptide. The resulting fusion peptide has the structural formula of formula XIV:

Xaai-Xaa2-Xaa3-Xaa4-Xaa5_XaE5-Xaa7-Xaa8-Xaa9-Xaai〇-

Xaai 1 -Xaai 2_Xaai 3_Xaai4_X (10) 15_Xaa! 6_Xaai 7-Xaai 8_Xaai 9_Χ3Β2〇·Χ New 21_

Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27_Xaa28-Xaa29-Xaa3〇 (SEQ ID 5 NO: 118), wherein:

Xaa!, X lean a4, Xaa5, Xaa8, Xaan, Xaa12, Xaai5, Xaai6, Xaais, Xaa!9, Xaa, 2, Xaa23, Xa&26, and Xaa3〇 are each a polar amino acid; and 10 Xaa2 , Xaa3, Xaa6, Xaa7, Xaa9, Xaa1G, Xaa13, Xaa14,

Xaai7, Xaa2〇, Xaa2i, Xaa24, Xaa25, Xaa27, and Xaa28 are each a non-polar amino acid. In several embodiments, the invention provides a sequence having one of the sequences corresponding to the amino acid N-terminal peptide of SEQ ID NO: 117, which is linked to one of the N-terminal peptides of the formula XI v peptide Fusion peptide. In another embodiment, the peptide of the present invention is a peptide comprising at least 14 contiguous amino acids of any of the foregoing peptides. In some embodiments, the non-polar amino acid is selected from the group consisting of: (1) alanine, valine, leucine, methionine, iso-amine 20 acid, benzene Alkalamine, and tryptophan; or (2) valine, leucine, isoleucine, phenylalanine, and tryptophan. In some embodiments, the polar amino acid is selected from the group consisting of arginine, aspartame, aspartic acid, cysteine, glutamic acid, glutamine, Histamine, homocysteine, lysine, hydroxy lysine, ornithine, serine and threonine; or (fantasy 34 200837075 aminic acid, aspartic acid, valine, cysteine Aminic acid and lysine. In another embodiment, the peptide of the present invention has arginine, cysteine, amylin, serine, valine, two aspartic acid, two An amino acid composition consisting of leucine, two isoleucine, and three tryptophan residues. For example, the peptide has an amino acid residue of SEq ID N〇: 92 or 1〇2. In another embodiment, the peptide of the present invention has arginine, cysteine, glutamic acid, two serines, valine, two aspartic acid, two leucines, two An amino10 acid composition comprising isoleucine and three tryptophan residues. For example, the peptide has the amino acid residue of SEQ ID NO: 93 or 101. In another embodiment, The peptide of the invention From arginine, cysteine, glutamic acid, two serines, valine, three aspartic acid, two leucines, two isoleucines and three tryptophan residues The amino group 15 acid composition consisting of. For example, the peptide has the amino acid residue of SEQ ID NO: 94 or 100. In another embodiment, the peptide of the present invention has arginine, cysteine Amino acid, glutamic acid, two serines, valine, three aspartic acid, two leucine, two isoleucine, three tryptophan and phenylalanine residues A composition of an amino acid composition. For example, the peptide has the amino acid residue of SEQ ID NO: 95 or 99. In another embodiment, the peptide of the present invention has arginine, cysteine , glutamic acid, two serines, valine, three aspartic acid, two leucine, two isoleucine, three tryptophan, phenylalanine and from 35 200837075 An amino acid composition consisting of a residue. For example, the peptide has the amino acid residues of SEQ ID NOS: 43 and 96-98. Another aspect of the invention comprises an excipient selected from SEQ ID NO: 43 And 91-102 a peptide of one of the amino acid sequences of the constituent group. In several 5 embodiments, the peptide is 14 to 50 D- or L-amino acids, and is selected from the group consisting of SEQ ID NO: 43 And an amino acid sequence of one of the groups consisting of 91-102, and the peptide has an amphipathic α-helical structure. Another aspect of the invention is an amine group having any one of SEQ ID NOS: 4-86 a peptide of an acid sequence. For example, the peptide has SEQ ID NO.·10 6 , 8, 12, 13, 14, 21, 23, 24, 27, 28, 30, 32, 37, 44, 47, 48 And any one of amino acid sequences of 53. Another aspect of the invention is having SEQ ID NO: 80, SEQ ID NO: 79 - SEQ ID NO: 91 ' SEQ ID NO: 97 ^ SEQ ID NO: 98, SEQ ID NO: 119, SEQ ID NO: 123, SEQ ID NO: 124, 15 SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 130, and SEQ ID NO: 130-144 A peptide of a base acid sequence. In one embodiment, the present invention provides an isolated peptide having from 14 to 50 D- or L-amino acids, wherein the peptide has antiviral activity or resistance against xanthrosis 20 orthopedic virus Human immunodeficiency virus antiviral activity, wherein the peptide has an amphipathic "_helix structure. In several embodiments, the amino acid of the peptide comprises a cysteine residue, which may be a serine acid The N-terminus can be located at four positions of the N-terminus of the serine acid. It can also be located at a position 11 relative to the 2^ end. 36 200837075 In another aspect, the present invention provides a method for deactivating a virus and a virus supplementation. Contacting any of the foregoing peptides, or contacting the mammal with any of the other pharmaceutical compositions or compositions. In another aspect, the invention provides for the prevention or treatment of a viral infection in a mammal. 5 The method of 'incorporating an effective amount of any of the foregoing peptides' into the (10) or administering to the mammal any of the above-mentioned pharmaceutics (four) or compositions. Other characteristics and advantages of the present invention Detailed Description section, and later by the scope of the patent applicant will be more apparent. Drawings briefly described 10

Figure 1 shows the position of the peptide relative to the HCV polyprotein genotype la (H77 isolate, with SEQ ID NO: 1} and the corresponding anti-hcv activity. 13 of the peptides tested were inhibited. Infectivity of 9% or more. Figure 2 shows peptide 1 (A) with amino acid sequence 15 column SWLRDIWDWICEVLSDFK (SEQ ID NO..43) with L-amino acid or D-amino acid. Initiation of HCV infection when added to the virus before exposure to the virus; (B) End of ongoing HCV infection; (C) inhibition of HCV infection by growing quiescent Huh-7 cells; (D) entry into the cell and E) Inhibition of intracellular HCV particle infectivity. As shown in two separate experiments, the EC5 of peptide 1 is approximately 300 nM 20 (F and G). Figure 3 is a histogram showing inhibition of Huh by various synthetic peptides. -7.5.1 HCV infection of cells in which peptide 1 (SEQ ID NO: 43) is the most potent inhibitor of viral infection. Figure 4A-G shows the inhibitory properties of peptide 1 (SEQ ID NO: 43) and 37 200837075 Information on stability. The L-isomer and D-isomer of peptide 1 are highly inhibitory (A). The peptide 1 acts on the virus (8) and destroys the hCV virion (D). It is viricidal (C) for HCV. The L-isomer and 〇-isomer are effective in membrane permeability (E). The D form of peptide 1 shows enhanced serum stability (?) and 5 is the lower IC5Q(G). Figure 5A-D shows the effective concentration (EC%) and toxicity (IC5〇) of the L-form and D· form of peptide 1 (SEQ ID NO: 43). Results. Figure 5A shows that the percentage of infected Huh-7.5.1 cells is a function of the concentration of the peptide 1 L-form, showing that the EQo of the peptide 1 L-form is 0.6 micromolar. Figure 5B shows the sense 10 The percentage of infected Η^_7·5·1 cells is a function of the concentration of the peptide 1 D-form (having D_amino acid instead of L. amino acid), showing that the peptide 1 D-form is 1.0 micron. Mohr concentration. Figure 5C shows that the percentage of viable Huh-7 cells is a function of the concentration of the L-form (square symbol) and D-form (diamond symbol) of peptide 1, showing the LC of the L-form of peptide 1. % is 46 μmol concentration and the LC% of the peptide it 15 D-form is 87 micromolar. Figure 5D shows the percentage of viable Huh-7.5.1 cells in the L-form of peptide 1 (square symbol) And the function of the concentration of the D-form (diamond symbol), showing victory The LC50 of the L-form of peptide 1 is 64 micromolar and the LC% of the D-form of peptide 1 is 105 micromolar. Figures 6A-E show two of peptide 1 (SEQ ID NO: 43). Affinity α-spiro 20 spin properties. The spiral wheel diagram of Figure 1 shows the distribution of the amino acid to obtain a hydrophilic (or polar) face and a water repellent (or non-polar) face (Fig. 6). The results of the circular two-color diagram show the L-isomer of peptide 1 and the α-helix structure of the D-isomer (Fig. 6), dimethylamino naphthalenesulfonate on the peptide 1 L-isomer and Effect of the D-isomer 〇1_ Helical structure (Fig. 6C), and α with a C-terminal truncation (Fig. 6D) and Ν 38 200837075 End truncation (Fig. 6E) Spiral structure. The sequences of these truncated peptides are provided in Table 9. Fig. 7 shows the results of the analysis of the liposome release assay obtained for the plurality of truncated mutants of peptide 1 (SEQ ID NO: 43). The sequences of these truncated peptides are presented in Table 9. Figure 8 is a line graph showing that peptide 1 (SEQ ID NO: 43; shown as #7208 in this icon) does not block vesicular stomatitis virus (VSV) infection. Additional studies indicate that peptide 1 does not block influenza virus (flu virus), bovine virus, Borna disease virus, lymphocytic choriomeningitis 10 or adenovirus infection. Figure 9 is a line graph showing the peptide 2022 (peptide 1) having the sequence SWLRDIWDWICEVLSDFK (SEQ ID NO. 43) and the peptide 2013 having the sequence SWLRDIWDWICEVL (SEQ ID NO: 92) as indicated by the ELISA. Inhibition of 100% dengue virus 15 staining. The peptide 2017 with the sequence LRDIWDWICEVLSDFK (SEQ ID NO.. 107) has a slightly lower activity and inhibits dengue virus infection by about 84%. Figure 10A-D is a line graph showing the percent inhibition of dengue virus infection by various peptides. Figure 10A shows dose dependence of dengue virus-infected peptide 2022 (peptide 20 1 ; SEQ ID NO: 43), peptide 2013 (SEQ ID NO: 92), and peptide 2017 (SEQ ID NO: 107) Inhibition, such as intracellular detection of dengue virus antigen staining cells by FACS analysis. As shown, at the concentration of 20 μΜ, almost 100% of the dengue virus infection was inhibited by the peptide 2022 (peptide 1) and the peptide 2013, as indicated by FACS. Peptide 2017 at 20μΜ has a slightly lower activity of 39,370,370,75, inhibiting dengue virus infection by up to approximately 8〇%. The first 〇B-D diagram also shows that the lentivirus mutant inhibits dengue virus infection, and the inhibitory effect of the amphiphilic peptide belonging to the mutant 1 mutant is verified, such as intracellular FACS staining. These data show that as long as the amphiphilic nature of the peptide structure is maintained, the variant 5 and homologue of peptide 1 retains inhibitory activity. Note that peptide 1 (SEQ ID NO: 43) is referred to as peptide L_72〇8 in Figure 10B-D. Figure 11 shows that peptide 1 (SEQ ID NO: 43; referred to as 2022 in this figure) and peptide 2012 (SWLRDIWDWICEVLSD, SEQ ID NO: 94) are associated with infectious virus 10 before virus is added to Huh-7 cells. The result of the ability of these peptides to inhibit West Nile virus (WNV) infection during culture. In this figure, the virus was added to the Huh-7 cells in the micro-valency plate, and the 102 to 1 PFU of WNV was pre-treated with DMSO (first column) or peptide 1 (18 micromolar concentration). Three-column) or peptide 2012 (fifth column) co-culture. The seventh column shows uninfected cells. After 5 days, the well plates were stained with WNV 15 specific antibodies. As shown, the viral antigen has been readily detected across the entire monolayer co-cultured with DMSO-treated virus. Conversely, there were only 4-5 staining cell focuss in cultures inoculated with 105 PFU of peptide-treated virus, whereas there was no evidence of infection in cultures vaccinated with lower doses of virus. Figure 12 is a graphical representation showing both the L-form and the D-form of peptide 1, i.e., 20 L-2022 [SWLRDIWDWICEVLSDFK] (SEQ ID NO: 43) and D-2022 (SEQ ID NO with D-amino acid: 43) HIV-1 infection can be inhibited at concentrations ranging from 1.25 micromolar to 5.0 micromolar. Also shown is that peptide 1818 (DIWDWICEVLSDFK) (SEQ ID NO: 108) is a N-terminal truncated 14-mer version of peptide 1 with similar antiviral activity; and peptide 40 200837075 2054 (SWLRDIWDWICEV) (SEQ ID NO: 103) The C-terminal truncated 13-mer analog of the peptide 1 has a slightly lower activity. Conversely, the peptide 2015 (SWLRDIWDWI) (SEQ ID NO: 105) is a C-terminal truncated 10-mer version of peptide 1 which is not active; peptide 6938 5 (LYGNEGCGWAGWLLSPRG) (SEQ ID NO: 6) is derived The 18-mer from the HCV core protein is also inactive. Figure 13A-B illustrates that L-7208 HS peptide (SIWRDWVDLICEFLSDWK, SEQ ID NO: 97) has activity nearly equal to highly active L-7208 SEQ ID NO: 43 peptide (also known as 10 L-2022 peptide or "peptide 1"). The L-7208 HS peptide has the same amino acid composition as the L-7208 peptide, but the water-reducing amino acid of the L-7208 HS peptide (SEQ ID NO. 97) has been broken up, thereby changing Amino acid sequence, but maintaining its amphiphilic nature. Thus, L-7208 HS peptide (SEQ ID NO: 97), 1-7208 peptide (SEQ ID NO: 43) and D-7208 peptide (SEQ ID NO: 43 15 contains D. amino acid instead of L -Amino acid) substantially inhibits 100% HIV infection at a concentration of about 20 micromolar. Another peptide (referred to as 3229, SWRLDIWDWICESVLDFK, SEQ ID NO: 119) whose amino acid is exchanged to reduce its amphiphilic properties, but which has the same amino acid as the L-7208 (SEQ ID NO: 43) peptide Composition, very little, if any, is active. This 20-specific data indicates that the amphipathic nature of the peptide is important for the activity, but the sequence of the peptide is not particularly limited. The results for HIVR9BaL from 293 T cells are shown in Figure 13A. The results obtained from CEM T cells for HIV R9BaL are shown in Figure 13B. Figures 14A-C show that the peptide is extracellularly stabilized against HIV-1 virions. Example 41 200837075 For example, Figure 14A shows that after treatment of HIV-1 preparation with peptide 1 (labeled as l-7208 (SEQ ID NO: 43)), the infectious virion releases a large amount of free HIV-1 shell. , indicating that the virion is dissolved by the peptide. Conversely, after treatment with HIV-1 virions in DMSO or with control peptide 6938 5 (LYGNEGCGWAGWLLSPRG) (SEQ ID NO: 6), the HIV-1 shell was not substantially released. Consistent with these observations, the number of shells associated with the virus treated with L-7208 (SEQ ID NO: 43) (Fig. 14B) was reduced, but treated with peptide 6938 (LYGNEGCGWAGWLLSPRG, SEQ ID NO: 6) The sample is no. Finally, Figure 14C shows HIV internalized into cells after treatment with 10 DMSO and 5 or 10 micromolar concentration peptide 6938 (SEQ ID NO: 6) or L-7208 (SEQ ID NO: 43). -1 Percentage of the housing. When cells were treated at 5 or 10 micromolar concentrations of 6938 (SEQ ID NO: 6), approximately 1% of the control number of HIV-1 capsids were internalized; via the use of peptide L-7208 (SEQ ID NO: 43) Treatment, the inhibition of 15 within the HIV-1 shell was 10 times. Figure 15 is a bar graph showing that the peptide having an amphiphilic structure similar to the amphipathic structure of peptide 1--7208 (SEQ ID NO: 43) can also strongly inhibit HIV infection. Thus, an amphiphilic peptide having potent anti-HIV activity includes peptide 3222 (SEQ ID NO: 127), peptide 3226 (SEQ ID NO: 128), 20 peptide 3228 (SEQ ID NO: 130), peptide L-7208 2D to 2 Pro (SEQ ID NO: 91) and L-7208 HS carry a dissociated hydrophilic amino acid (KWLCRIWSWISDVLDDFE, SEQ ID NO.. 98). Figure 16A-D: peptide 1 (SEQ ID NO: 43; amphipathic "viral agent") not only neutralizes cell-free HIV, but also neutralizes cells and binds to HIV 2008 and 2007. (A) CD4+ T-lymphocytes, macrophages or DCs (0.1 x 16 cells) were exposed to NL4.3 BaL (1 Ng ρ24) 1 曰, washed three times with the culture medium, and cultured in a flat-bottom 96-well plate. Wild type peptide 1 or its non-amphibious variant (SEQ ID NO: 119; 5 μΜ) was added to cd4+ 5 Τ-cell, python and DC (Figs. 1 to 3) with virus before Τ cell culture Add to DCs pulsed with HIV (Figure 4) or to T cells 3 weeks after infection (Figure 5). Supernatants were collected at different days and the virus was monitored by p24 ELISA. The error bars represent the standard deviation of the replicates. These experiments represent three independent experiments using three different donors. (B) 293 T cells infected with NL4.3 or NL4.3 BaL for 24 hours were transfected with a peptide (5 μM) at 37 ° C or without using a peptide for 1 hour, and the peptide was removed by washing. The infectivity of the virus released at 293 ΤΖΜ on the sputum cell score 24 hours after the peptide treatment. The infection was determined by galactosidase activity 48 hours after infection. The error bars represent the standard deviation of the repeated tests. These experiments represent two independent experiments. (C) TZM cells were exposed to pNL4.3-AEnv virus (1 Nike p24) of VSVG pseudotyped by gpl60NL4.3 (X4), gpl60 BaL (R5) or with or without peptide (5 μM), respectively. The infection was determined by point-galactosidase activity 48 hours after infection. The error bars represent the standard deviation of the replicates. Data are expressed as a percentage of infection. (D) TZM cells were pretreated with peptide (5 μM) for 1 hour, 2 hours, 4 hours, and 8 hours, after thorough 20 washes to remove the peptide, and then exposed to NL4.3 (1 Nike Ρ 24); or cell first Exposure to virus and addition of peptides after 1, 2, 4 and 8 hours. The infection was determined by galactosidase activity 48 hours after infection. The error bars represent the standard deviation of the repeated experiments. Section 17 - Map: peptide 1 (SEQ ID NO.·43; amphipathic "viricide 43 200837075") destroys the membrane core protein of HIV and the core protein of the shell intact. (a) Purified NL4.3 virus (20 ng p24 in PBS) with or without peptide 1 (5 μM) was incubated at 37 ° C for 30 minutes and loaded on a 20-70% sucrose gradient. The collected gradients were analyzed for the shell (p24 ELISA and immune dots), 5 RT (by exoRT assay), and gp41 content (by immunoblots). The selected density (g/cm3) shown in the lower two figures is determined by measuring the refractive index. (B) For virus attachment, TZM cells (500,000) were exposed to 1 ng of p24 of NL4.3 for 1 hour at 4 °C, thoroughly washed to remove unbound virus and dissolved. For virus internalization, cells were exposed to virus for 2 hours at 37 ° C, washed, and treated with trypsin to remove the attached virus and dissolve. The amount of virus attached to the internalization virus was determined by cell lysate by P24 ELISA. The error bars indicate the standard deviation of the repeated tests. The data is expressed as a percentage of attachment or a percentage of internalization. These experiments represent two independent experiments. (C) Same as (A), but the virus is treated with a decreasing concentration of peptide 1 at 37 °C for 3 minutes (top left); treated with 5 μΜ 15 peptide 1 at 37 ° C for 15, 30 and 60 minutes (bottom left) Figure); treated at 4 ° C, 25 ° C or 37 ° C for 30 minutes (top right); and treated at pH 8, 7, 6 and 5 for 30 minutes (bottom right). Gradient selection was performed by P24 ELISA to analyze HIV shells. (D) Same as (A), but the virus is treated with peptide 1 or with its non-amphibious variant (SEQ ID NO: 119). (E) Same as (A), but the virus was first trypsinized at 37 ° C for 15 minutes and 20 minutes in 10% FCS to neutralize trypsin, microcentrifuged at 4 ° C for 90 minutes 'resuspend again and immediately load to The sucrose gradient was used to assess viral integrity by p24 ELISA. Figure 18A-F: peptide 1 (SEQ ID NO: 43; amphipathic "viral agent") inhibits HIV reproductive epithelial migration and LC/DC forwarding of HIV. (A) No 44 200837075 Cell-containing HIV or cell-bound mv at 37 It is added to the top surface of a primary reproductive epithelial cell (PGEC) for 8 hours, and the amount of virus transported through the cell is the lower chamber corresponding to the bottom surface of the PGEC. Quantification of p24 ELISA. In order to determine the effect of peptide 1 (SEQ ID NO: 43) on HIV translocation, just add the peptide 1 after the virus was added with 5 spGEC, and the effect of peptide 1 and its non-amphipathic variant (SEQ ID NO ··119) Comparison. The results are expressed as a percentage of p24 originally inoculated. The error bars represent the standard deviation of the replicates. The results were representatives of 4 independent experiments using PGEC derived from 4 donors. (B) PGEC^ was treated twice with 200 μΜ peptide 1 (SEQ ID NO ··43) or 0.01% saponin for 10 weeks. No washing was done to maintain continuous exposure of the cells to the peptide. After overnight incubation, cell quantitation (CellQuanti)-MTTTM reagent was added and cell viability was quantified by OD 570 nm reading. As for the 18 C-F map, the epidermal sheet was infected with HIV NL4.3-BaL-eGFP (100 ng p24) and co-cultured directly with ΙΟμΜ peptide 1 (SEQ ID NO: 43) or the DMSO control group. After 3 ,, 15 remove the epidermal sheet and add 200,000 CCR5 + Jurkat cells to 4 曰. The migrated DC/LC epidermal cells (Day 3) and co-cultured samples (Sections 5 and 7) were analyzed by FACS for green fluorescent protein (GFP) expression. (C) Migration of DC/LC HIV infection is expressed as a percentage of total cells. The error bars represent the standard deviation of the replicates. (D) Co-cultivation Further on the 7th day, FACS was used to analyze the infection. Shows the percentage of infected cells. (E) The donor variation rate shown in Section 5. The error bars represent the standard deviation of the replicates. (F) DC (50,000) exposed to HIV-1 NL4.3-eGFP (X4), NL4.3-BaL-eGFP (R5) at 37 °C or exposed to NL4.3 gpl60 env (25 Ng p24) Pseudotypes of NL4.3 Δ Env-eGFP together with 1 〇μΜ peptide 1 or DMSO control group for 2 hours. Cell washing 45 200837075 Three times, activated CD4+ T cells were added for 3 days, and GFP expression was measured by FACS. The error bars represent the standard deviation of the replicates. These results represent 3 independent experiments. C Embodiment 3 5 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention relates to a peptide which prevents or treats a viral infection or which can be used to activate a virus before it enters a cell. The present invention relates to the discovery of certain peptides derived from the hepatitis C virus polyprotein, for example, a peptide having the sequence of SEQ ID NO: 4-61 can prevent or treat a mammal from being reduced by several classes of viruses. Pages of the disease, aphrodisiac virus, respiratory syncytial virus (RSV) and HIV, as well as the ability to deactivate these viruses. The present invention also relates to the discovery that several peptides derived from the HCV polyprotein (SEQ ID NO: 1) are highly effective for inhibiting the infection of the ¥ and the infection of the Flaviviridae virus. In addition, the present invention relates to the discovery that "peptide pI" 15 (SEQ ID NO: 43) derived from the NS5A membrane anchoring functional site (NS5A-1975) is particularly potent against HIV, hepatitis C virus, aphrodisiac virus, RSV and Flaviviruses such as dengue virus and West Nile virus. For example, 20μ Μ peptide 1 completely inhibits HIV infection, and a low 〇 3 μ Μ concentration can strongly inhibit HIV infection. In addition, the peptide is useful for the prevention or treatment of infections in a variety of HIV strains, including HIV germline infections using CXCR4 and CCR5 as accessory receptors. The peptide of the present invention includes, for example, a peptide having the sequences set forth in SEQ ID NOS: 4-61, 91·102, 79-80, 119, 123-128, and 130-144, and which can form an α-helical structure and A peptide of from about 8 to about 50 amino acids which inhibits viral infection in mammalian cells. The present invention provides an antiviral peptide or an anti-virulence peptide composition, a plurality of compositions and compositions comprising the antiviral peptides, and a method of inhibiting viral infection of mammalian cells using such peptides. The invention also provides a manufactured article comprising such antiviral peptides. According to the present invention, the peptide of the hepatitis C virus polyprotein derived from the hepatitis C genotype la (H77) and the hepatitis genotypes IB, 2A, 4A, 5A and 6A has a strong peptide. Inhibition of HIV infection. Thus, for example, the HCV polyprotein sequence from which the peptide was originally obtained has the sequence seQIDNO: 1, which is available from the NCBI database under access number NP 671491 (gi: 22129793). This HCV poly egg 10 white amino acid sequence (SEQ ID NO: 1) is as follows. 15

20 25

1 MSTNPKPQRK TKRNTNRRPE DVKFPGGGQI VGGVYLLPRR 41 GPRLGVRTTR KTSERSQPRG RRQPIPKDRR STGKAWGKPG 81 RPWPLYGNEG LGWAGWLLSP RGSRPSWGPT DPRHRSRNVG 121 KVIDTLTCGF ADLMGYIPVV GAPLSGAARA VAHGVRVLED 161 GVNYATGNLP GFPFSIFLLA LLSCITVPVS AAQVKNTSSS 201 YMVTNDCSND SITWQLEAAV LHVPGCVPCE RVGNTSRCWV 241 PVSPNMAVRQ PGALTQGLRT HIDMVVMSAT FCSALYVGDL 281 CGGVMLAAQV FIVSPQYHWF VQECNCSIYP GTITGHRMAW 321 DMMMNWSPTA TMILAYVMRV PEVIIDIVSG AHWGVMFGLA 361 YFSMQGAWAK VIVILLLAAG VDAGTTTVGG AVARSTNVIA 401 GVFSHGPQQN IQLINTNGSW HINRTALNCN DSLNTGFLAA 441 LFYTNRFNSS GCPGRLSACR NIEAFRIGWG TLQYEDNVTN 481 PEDMRPYCWH YPPKPCGVVP ARSVCGPVYC FTPSPVVVGT 521 TDRRGVPTYT WGENETDVFL LNSTRPPQGS WFGCTWMNST 561 GFTKTCGAPP CRTRADFNAS TDLLCPTDCF RKHPDATYIK 601 CGSGPWLTPK CLVHYPYRLW HYPCTVNFT station FKIRMYVGGV 641 EHRLTAACNF TRGDRCDLED RDRSQLSPLL HSTTEWAILP 681 CTYSDLPALS TGLLHLHQNI VDVQYMYGLS PAITKYVVRW 721 EWVVLLFLLL ADARVCACLW MLILLGQAEA ALEKLVVLHA 761 ASAANCHGLL YFAIFFVAAW HIRGRVVPLT TYCLTGLWPF 801 CLLLMALPRQ AYAYDAPVHG QIGVGLLILI TLFTL TPGYK 841 TLLGQCLWWL CYLLTLGEAM Work QEWVPPMQV RGGRDGIAWA 47 30 200837075

881 VTIFCPGVVF DITKWLLALL GPAYLLRAAL THVPYFVRAH 921 ALIRVCALVK QLAGGRYVQV ALLALGRWTG TYIYDHLTPM 961 SDWAASGLRD LAVAVEP 工 F SPMEKKVIVW GAETAACGDI 1001 LHGLPVSARL GQEILLGPAD GYTSKGWKLL APITAYAQQT

5 1041 RGLLGAIVVS MTGRDRTEQA GEVQILSTVS QSFLGTTISG

1081 VLWTVYHGAG NKTLAGLRGP VTQMYSSAEG DLVGWPSPPG

1121 TKSLEPCKCG AVDLYLVTRN ADVIPARRRG DKRGALLSPR

1161 PISTLKGSSG GPVLCPRGHV VGLFRAAVCS RGVAKSIDFI

1201 PVETLDVVTR SPTFSDNSTP PAVPQTYQVG YLHAPTGSGK

10 1241 STKVPVAYAA QGYKVLVLNP SVAATLGFGA YLSKAHGINP

1281 NIRTGVRTVM TGEAITYSTY GKFLADGGCA SGAYD Engineering CD

1321 ECHAVDATSI LGIGTVLDQA ETAGVRLTVL ATATPPGSVT

1361 TPHPDIEEVG LGREGEIPFY GRAIPLSCIK GGRHLIFCHS

1401 KKKCDELAAA LRGMGLNAVA YYRGLDVS 工 PAQGDVVVVA

15 1441 TDALMTGYTG DFDSVIDCNV AVTQAVDFSL DPTFTITTQT

1481 VPQDAVSRSQ RRGRTGRGRQ GTYRYVSTGE RASGMFDSVV

1521 LCECYDAGAA WYDLTPAETT VRLRAYFNTP GLPVCQDHLE

1561 FWEAVFTGLT HIDAHFLSQT KQAGENFAYL VAYQATVCAR

1601 AKAPPPSWDA MWKCLARLKP TLAGPTPLLY RLGPITNEVT

20 1641 LTHPGTKYIA TCMQADLEVM TSTWVLAGGV LAAVAAYCLA

1681 TGCVSIIGRL HVNQRVVVAP DKEVLYEAFD EMEECASRAA

1721 LIEEGQRIAE MLKSKIQGLL QQASKQAQDI QPAMQASWPK

1761 VEQFWARHMW NFISGIQYLA GLSTLPGNPA VASMMAFSAA

1801 LTSPLSTSTT LLNIMGGWL ASQIAPPAGA TGFVVSGLVG

25 1841 AAVGSIGLGK VLVDILAGYG AGISGALVAF KIMSGEKPSM

1881 EDVINLLPGI LSPGALVVGV ICAAILRRHV GPGEGAVQWM

1921 NRLIAFASRG NHVAPTHYVT ESDASQRVTQ LLGSLTITSL

1961 LRRLHNWITE DCPIPCSGSW LRDVWDWVCT ILTDFKNWLT

2001 SKLFPKLPGL PFISCQKGYK GVWAGTGIMT TRCPCGANIS

30 2041 GNVRLGSMRI TGPKTCMNTW QGTFPINCYT EGQCAPKPPT

2081 NYKTAIWRVA ASEYAEVTQH GSYSYVTGLT TDNLKIPCQL

2121 PSPEFFSWVD GVQIHRFAPT PKPFFRDEVS FCVGLNSYAV

2161 GSQLPCEPEP DADVLRSMLT DPPHITAETA ARRLARGSPP

2201 SEASSSVSQL SAPSLRATCT THSNTYDVDM VDANLLMEGG

35 2241 VAQTEPESRV PVLDFLEPMA EEESDLEPSI PSECMLPRSG

2281 FPRALPAWAR PDYNPPLVES WRRPDYQPPT VAGCALPPPK

2321 KAPTPPPRRR RTVGLSESTI SEALQQLAIK TFGQPPSSGD

2361 AGSSTGAGAA ESGGPTSPGE PAPSETGSAS SMPPLEGEPG

2401 DPDLESDQVE LQPPPQGGGV APGSGSGSWS TCSEEDDTTV 48 200837075 10

15

2441 CCSMSYSWTG ALITPCSPEE EKLPINPLSN SLLRYHNKVY 2481 CTTSKSASQR AKKVTFDRTQ VLDAHYDSVL KDIKLAASKV 2521 SARLLTLEEA CQLTPPHSAR SKYGFGAKEV RSLSGRAVNH 2561 IKSVWKDLLE DPQTPIPTTI MAKNEVFCVD PAKGGKKPAR 2601 LIVYPDLGVR VCEKMALYD work TQKLPQAVMG ASYGFQYSPA 2641 QRVEYLLKAW AEKKDPMGFS YDTRCFDSTV TERDIRTEES 2681 lYQACSLPEE ARTAIHSLTE RLYVGGPMFN SKGQTCGYRR 2721 CRASGVLTTS MGNTITCYVK ALAACKAAGI VAPTMLVCGD 2761 DLVVISESQG TEEDERNLRA FTEAMTRYSA PPGDPPRPEY 2801 DLELITSCSS NVSVALGPRG RRRYYLTRDP TTPLARAAWE 2841 TVRHSPINSW LGNIIQYAPT station WVRMVLMTH FFSILMVQDT 2881 LDQNLNFEMY GSVYSVNPLD LPAIIERLHG LDAFSMHTYS 2921 HHELTRVASA LRKLGAPPLR VWKSRARAVR ASLISRGGKA 2961 AVCGRYLFNW AVKTKLKLTP LPEARLLDLS SWFTVGAGGG 3001 DIFHSVSRAR PRSLLFGLLL LFVGVGLFLL PAR can be used as another example of the amino acid sequence of the polyprotein of the HCV-derived peptides related to NCBI database Available with access number BAB32872 (gi: 13122262). Reference Kato et al., J. Med. Virol. 64: 334-339 (2001). Such an HCV system is isolated from a patient with a swollen hepatitis, and its amino acid sequence (SEQ ID NO: 2) is as follows. 1 41 81 121 25 161 201 241 281 321 30 361 401 441 481 521

MSTNPKPQRK

GPRLGVRTTR

RPWPLYGNEG

KVIDTLTCGF

GVNYATGNLP

YMVTNDCSND

PVSPNMAVRQ

CGGVMLAAQV

DMMMNWSPTA

YFSMQGAWAK

GVFSHGPQQN

LFYTNRFNSS

PEDMRPYCWH

TDRRGVPTYT

TKRNTNRRPE KTSERSQPRG LGWAGWLLSP ADLMGYIPVV GFPFS Labor FLLA SITWQLEAAV PGALTQGLRT FIVSPQYHWF TMILAYVMRV VIVILLLAAG IQLINTNGSW GCPGRLSACR YPPKPCGVVP WGENETDVFL

DVKFPGGGQI

RRQPIPKDRR

RGSRPSWGPT

GAPLSGAARA

LLSCITVPVS

LHVPGCVPCE

HIDMVVMSAT

VQECNCSIYP

PEVIIDIVSG

VDAGTTTVGG

HINRTALNCN

NIEAFRIGWG

ARSVCGPVYC

LNSTRPPQGS

VGGVYLLPRR

STGKAWGKPG

DPRHRSRNVG

VAHGVRVLED

AAQVKNTSSS

RVGNTSRCWV

FCSALYVGDL

GTITGHRMAW

AHWGVMFGLA

AVARSTNVIA

DSLNTGFLAA

TLQYEDNVTN

FTPSPVVVGT

WFGCTWMNST 49 200837075

561 GFTKTCGAPP CRTRADFNAS TDLLCPTDCF RKHPDATYIK

601 CGSGPWLTPK CLVHYPYRLW HYPCTVNFTI FKIRMYVGGV

641 EHRLTAACNF TRGDRCDLED RDRSQLSPLL HSTTEWAILP

681 CTYSDLPALS TGLLHLHQNI VDVQYMYGLS PAITKYWRW

5 721 EWVVLLFLLL ADARVCACLW MLILLGQAEA ALEKLVVLHA

761 ASAANCHGLL YFAIFFVAAW HIRGRVVPLT TYCLTGLWPF

801 CLLLMALPRQ AYAYDAPVHG QIGVGLLILI TLFTLTPGYK

841 TLLGQCLWWL CYLLTLGEAM IQEWVPPMQV RGGRDGIAWA

881 VTIFCPGVVF DITKWLLALL GPAYLLRAAL THVPYFVRAH

10 921 ALIRVCALVK QLAGGRYVQV ALLALGRWTG TYIYDHLTPM

961 SDWAASGLRD LAVAVEPIIF SPMEKKVIVW GAETAACGDI 1001 LHGLPVSARL GQEILLGPAD GYTSKGWKLL APITAYAQQT

1041 RGLLGAIVVS MTGRDRTEQA GEVQILSTVS QSFLGTTISG

1081 VLWTVYHGAG NKTLAGLRGP VTQMYSSAEG DLVGWPSPPG

15 1121 TKSLEPCKCG AVDLYLVTRN ADVIPARRRG DKRGALLSPR

1161 PISTLKGSSG GPVLCPRGHV VGLFRAAVCS RGVAKSIDFI

1201 PVETLDVVTR SPTFSDNSTP PAVPQTYQVG YLHAPTGSGK

1241 STKVPVAYAA QGYKVLVLNP SVAATLGFGA YLSKAHGINP

1281 NIRTGVRTVM TGEAITYSTY GKFLADGGCA SGAYDIIICD

20 1321 ECHAVDATSI LGIGTVLDQA ETAGVRLTVL ATATPPGSVT

1361 TPHPDIEEVG LGREGEIPFY GRAIPLSCIK GGRHLIFCHS

1401 KKKCDELAAA LRGMGLNAVA YYRGLDVSII PAQGDVVVVA

1441 TDALMTGYTG DFDSVIDCNV AVTQAVDFSL DPTFTITTQT

1481 VPQDAVSRSQ RRGRTGRGRQ GTYRYVSTGE RASGMFDSVV

25 1521 LCECYDAGAA WYDLTPAETT VRLRAYFNTP GLPVCQDHLE

1561 FWEAVFTGLT HIDAHFLSQT KQAGENFAYL VAYQATVCAR

1601 AKAPPPSWDA MWKCLARLKP TLAGPTPLLY RLGPITNEVT

1641 LTHPGTKYIA TCMQADLEVM TSTWVLAGGV LAAVAAYCLA

1681 TGCVSIIGRL HVNQRVVVAP DKEVLYEAFD EMEECASRAA

30 1721 LIEEGQRIAE MLKSKIQGLL QQASKQAQDI QPAMQASWPK

1761 VEQFWARHMW NFISGIQYLA GLSTLPGNPA VASMMAFSAA

1801 LTSPLSTSTT ILLNIMGGWL ASQIAPPAGA TGFVVSGLVG

1841 AAVGSIGLGK VLVDILAGYG AGISGALVAF KIMSGEKPSM

1881 EDVINLLPG LSPGALVVGV WORK CAAILRRHV GPGEGAVQWM

35 1921 NRLIAFASRG NHVAPTHYVT ESDASQRVTQ LLGSLTITSL

1961 LRRLHNWITE DCPIPCSGSW LRDVWDWVCT ILTDFKNWLT

2001 SKLFPKLPGL PFISCQKGYK GVWAGTGIMT TRCPCGANIS

2041 GNVRLGSMRI TGPKTCMNTW QGTFPINCYT EGQCAPKPPT

2081 NYKTAIWRVA ASEYAEVTQH GSYSYVTGLT TDNLKIPCQL 50 200837075 5 10

15 20

2121 PSPEFFSWVD GVQIHRFAPT PKPFFRDEVS FCVGLNSYAV 2161 GSQLPCEPEP DADVLRSMLT DPPHITAETA ARRLARGSPP 2201 SEASSSVSQL SAPSLRATCT THSNTYDVDM VDANLLMEGG 2241 VAQTEPESRV PVLDFLEPMA EEESDLEPSI PSECMLPRSG 2281 FPRALPAWAR PDYNPPLVES WRRPDYQPPT VAGCALPPPK 2321 KAPTPPPRRR RTVGLSESTI SEALQQLAIK TFGQPPSSGD 2361 AGSSTGAGAA ESGGPTSPGE PAPSETGSAS SMPPLEGEPG 2401 DPDLESDQVE LQPPPQGGGV APGSGSGSWS TCSEEDDTTV 2441 CCSMSYSWTG ALITPCSPEE EKLPINPLSN SLLRYHNKVY 2481 CTTSKSASQR AKKVTFDRTQ VLDAHYDSVL KDIKLAASKV 2521 SARLLTLEEA CQLTPPHSAR SKYGFGAKEV RSLSGRAVNH 2561 workers KSVWKDLLE DPQTPIPTTI MAKNEVFCVD PAKGGKKPAR 2601 LIVYPDLGVR VCEKMALYDI TQKLPQAVMG ASYGFQYSPA 2641 QRVEYLLKAW AEKKDPMGFS YDTRCFDSTV TERDIRTEES 2681 IYQACSLPEE ARTAIHSLTE RLYVGGPMFN SKGQTCGYRR 2721 CRASGVLTTS MGNTITCYVK ALAACKAAG work VAPTMLVCGD 2761 DLVVISESQG TEEDERNLRA FTEAMTRYSA PPGDPPRPEY 2801 DLELITSCSS NVSVALGPRG RRRYYLTRDP TTPLARAAWE 2841 TVRHSPINSW LGN workers work QYAPI * IWVRMVLMTH FFSILMVQDT 2881 LDQNLNFEMY GSVYSVNPLD LPAIIERLHG LDAFSMHTYS 2921 HHELTR VASA LRKLGAPPLR VWKSRARAVR ASLISRGGKA 2961 AVCGRYLFNW AVKTKLKLTP LPEARLLDLS SWFTVGAGGG 3001 DIFHSVSRAR PRSLLFGLLL LFVGVGLFLL PAR Another 25 examples of HCV polyprotein amino acid sequences that can be used as a source of related peptides in the NCBI database with access number Q9WMX2匕丨:68565847) Acquired. See ncbi.nlm.nih.gov. This sequence was obtained from HCV Coni isolated strains. The amino acid sequence (SEQ ID NO.. 3) is as follows.

1 MSTNPKPQRK TKRNTNRRPQ DVKFPGGGQI VGGVYLLPRR 41 GPRLGVRATR KTSERSQPRG RRQPIPKARQ PEGRAWAQPG 81 YPWPLYGNEG LGWAGWLLSP RGSRPSWGPT DPRRRSRNLG 121 KVIDTLTCGF ADLMGYIPLV GAPLGGAARA LAHGVRVLED 161 GVNYATGNLP GCSFSIFLLA LLSCLTIPAS AYEVRNVSGV 201 YHVTNDCSNA SIVYEAADMI MHTPGCVPCV RENNSSRCWV 241 ALTPTLAARN ASVPTTTIRR HVDLLVGAAA LCSAMYVGDL 281 CGSVFLVAQL FTFSPRRHET VQDCNCSIYP GHVTGHRMAW 51 200837075 5 10 15 20 25 30

321 DMMMNWSPTA ALVVSQLLR station PQAVVDMVAG AHWGVLAGLA 361 YYSMVGNWAK VLIVMLLFAG VDGGTYVTGG TMAKNTLGIT 401 SLFSPGSSQK IQLVNTNGSW HINRTALNCN DSLNTGFLAA 441 LFYVHKFNSS GCPERMASCS PIDAFAQGWG PITYNESHSS 481 DQRPYCWHYA PRPCGIVPAA QVCGPVYCFT PSPVVVGTTD 521 RFGVPTYSWG ENETDVLLLN NTRPPQGNWF GCTWMNSTGF 561 TKTCGGPPCN station GGIGNKTLT CPTDCFRKHP EATYTKCGSG 601 PWLTPRCLVH YPYRLWHYPC TVNFTIFKVR MYVGGVEHRL 641 EAACNWTRGE RCNLEDRDRS ELSPLLLSTT EWQVLPCSFT 681 TLPALSTGLI HLHQNVVDVQ YLYGIGSAVV SFAIKWEYVL 721 LLFLLLADAR VCACLWMMLL IAQAEAALEN LVVLNAASVA 761 GAHGILSFLV FFCAAWYIKG RLVPGAAYAL YGVWPLLLLL 801 LALPPRAYAM DREMAASCGG AVFVGLILLT LSPHYKLFLA 841 RLIWWLQYF workers TRAEAHLQVW work PPLNVRGGR DAVILLTCAI 881 HPELIFTITK ILLAILGPLM VLQAGITKVP YFVRAHGLIR 921 ACMLVRKVAG GHYVQMALMK LAALTGTYVY DHLTPLRDWA 961 HAGLRDLAVA VEPVVFSDME TKVITWGADT AACGDIILGL 1001 PVSARRGREI HLGPADSLEG QGWRLLAPIT AYSQQTRGLL 1041 GCIITSLTGR DRNQVEGEVQ VVSTATQSFL ATCVNGVCWT 1081 VYHGAGSKTL AGPKGPITQM YTNVDQDLVG WQAPPGARSL 1121 TPCTCGSSDL YLVTRHADVI PV RRRGDSRG SLLSPRPVSY 1161 LKGSSGGPLL CPSGHAVGIF RAAVCTRGVA KAVDFVPVES 1201 METTMRSPVF TDNSSPPAVP QTFQVAHLHA PTGSGKSTKV 1241 PAAYAAQGYK VLVLNPSVAA TLGFGAYMSK AHGIDPNIRT 1281 GVRTITTGAP station TYSTYGKFL ADGGCSGGAY DIIICDECHS 1321 TDSTTILGIG TVLDQAETAG ARLVVLATAT PPGSVTVPHP 1361 ISilEEVALSST GEIPFYGKAI PIETIKGGRH LIFCHSKKKC 1401 DELAAKLSGL GLNAVAYYRG LDVSVIPTSG DVIVVATDAL 1441 MTGFTGDFDS VIDCNTCVTQ TVDFSLDPTF TIETTTVPQD 1481 AVSRSQRRGR TGRGRMGIYR FVTPGERPSG MFDSSVLCEC 1521 YDAGCAWYEL TPAETSVRLR AYLNTPGLPV CQDHLEFWES 1561 VFTGLTHIDA HFLSQTKQAG DNFPYLVAYQ ATVCARAQAP 1601 PPSWDQMWKC LIRLKPTLHG PTPLLYRLGA VQNEVTTTHP 1641 ITKYIMACMS ADLEVVTSTW VLVGGVLAAL AAYCLTTGSV 1681 VIVGRIILSG KPAIIPDREV LYREFDEMEE CASHLPYIEQ 1721 GMQLAEQFKQ KAIGLLQTAT KQAEAAAPVV ESKWRTLEAF 1761 WAKHMWNFIS GIQYLAGLST LPGNPAIASL MAFTASITSP 1801 LTTQHTLLFN ILGGWVAAQL APPSAASAFV GAGIAGAAVG 1841 SIGLGKVLVD work LAGYGAGVA GALVAFKVMS GEMPSTEDLV

52 35 200837075 10

15 20

1881 NLLPAILSPG ALVVGVVCAA ILRRHVGPGE GAVQWMNRLI 1921 AFASRGNHVS PTHYVPESDA AARVTQILSS LTITQLLKRL 1961 HQWINEDCST PCSGSWLRDV WDWICTVLTD FKTWLQSKLL 2001 PRLPGVPFFS CQRGYKGVWR GDGIMQTTCP CGAQITGHVK 2041 NGSMRIVGPR TCSNTWHGTF PINAYTTGPC TPSPAPNYSR 2081 ALWRVAAEEY VEVTRVGDFH YVTGMTTDNV KCPCQVPAPE 2121 FFTEVDGVRL HRYAPACKPL LREEVTFLVG LNQYLVGSQL 2161 PCEPEPDVAV LTSMLTDPSH ITAETAKRRL ARGSPPSLAS 2201 SSASQLSAPS LKATCTTRHD SPDADLIEAN LLWRQEMGGN 2241 workers TRVESENKV VILDSFEPLQ AEEDEREVSV PAEILRRSRK 2281 FPRAMPIWAR PDYNPPLLES WKDPDYVPPV VHGCPLPPAK 2321 APPIPPPRRK RTVVLSESTV SSALAELATK TFGSSESSAV 2361 DSGTATASPD QPSDDGDAGS DVESYSSMPP LEGEPGDPDL 2401 SDGSWSTVSE EASEDVVCCS MSYTWTGALI TPCAAEETKL 2441 PINALSNSLL RHHNLVYATT SRSASLRQKK VTFDRLQVLD 2481 DHYRDVLKEM KAKASTVKAK LLSVEEACKL TPPHSARSKF 2521 GYGAKDVRNL SSKAVNHIRS VWKDLLEDTE TPIDTTIMAK 2561 NEVFCVQPEK GGRKPARLIV FPDLGVRVCE KMALYDVVST 2601 LPQAVMGSSY GFQYSPGQRV EFLVNAWKAK KCPMGFAYDT 2641 RCFDSTVTEN DIRVEESIYQ CCDLAPEARQ AIRSLTERLY 2681 work GGPLTNSKG QNCGYRRCRA SGVLTTSCGN TLTCYLKAAA 2721 ACRAAKLQDC TMLVCGDDLV VICESAGTQE DEASLRAFTE 2761 AMTRYSAPPG DPPKPEYDLE LITSCSSNVS VAHDASGKRV 2801 YYLTRDPTTP LARAAWETAR HTPVNSWLGN work station MYAPTLWA 2841 RMILMTHFFS LLAQEQLEK ALDCQIYGAC YSIEPLDLPQ 2881 work station working QRLHGLSA FSLHSYSPGE INRVASCLRK LGVPPLRVWR 2921 HRARSVRARL LSQGGRAATC GKYLFNWAVR TKLKLTPIPA 2961 ASQLDLSSWF VAGYSGGDIY HSLSRARPRW FMWCLLLLSV 3001 GVGIYLLPNR 3〇 HCV can be obtained Additional examples of polyprotein sequences are available as other sources of antiviral peptides. For example, the hepatitis C virus Taiwan isolate can be obtained from the NCBI database under access number P29846 (gi: 266821). Reference ncbi.nlm.nih.gov. In infected cells, HCV polyproteins are cleaved by cellular protease 35 and viral proteases at multiple locations to produce structural and non-structural (NS) proteins. 53 200837075 The production of mature non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A and NS5B) is affected by two viral proteases. The first one has not been clearly characterized, and is cleaved at the NS2-NS3 linker; the second is a serine protease (hereinafter referred to as NS3 protease) contained in the N-terminal region of NS3, and the enzyme is subsequently cleaved by all of the downstream 5 of NS3. These include cis cleavage at the NS3-NS4A cleavage site, and trans cleavage of the remaining NS4A-NS4B, NS4B-NS5A, NS5A-NS5B positions. The NS4A protein is clearly useful as a cofactor for NS3 protease and may assist in membrane localization of NS3 and other viral replicase components. The formation of a complex of NS3 protease and NS4A is clearly required for the treatment of event 10, which increases the efficiency of protein decomposition at all locations. The NS3 protein also has nucleoside triphosphatase activity and RNA helicase activity. NS5B is an RNA-dependent RNA polymerase involved in HCV replication. The putative H C V non-structural (N S) protein provides the catalytic machinery necessary for viral replication. 181 amino acid before NS3 (residue I5 of the viral polyprotein shows a functional part of the NS3-containing serine protein plum, which treats all four downstream positions of the HCV polyprotein (C. Lin et al., j. Virol. 68, 8147-8157 (1994)) 〇HCV has three structural proteins, namely the N-terminal nucleocapsid protein (named "core") and two envelope glycoproteins, namely "E1" (also known as E) and rE2" 20 (also known as E2/NS1). Refer to Houghton et al., Hepatology 14:381-388' for a discussion of HCV proteins including E1 and E2. The E1 protein was tested to belong to the 32-35 kDa class and was converted to approximately 18 kDa. A single internal H-sensitive band. Conversely, E2 exhibits complex patterns consistent with the production of multiple classes when immunoprecipitated (Grakoui, ed. (1993) J. Virol 67: 1385-1395; Tomei 54 200837075 et al. (1993) J. Virol 67: 4017-4026). HCV envelope glycoproteins El and E2 form a co-precipitable stable complex (Grakoui et al. (1993) J. Virol. 67: 1385-1395; Lanford et al. (1993) Learning 197: 225-235;

Ralston et al. (1993) J. Virol 67:6753-6761) 〇 5 Antiviral peptides One aspect of the present invention is an antiviral peptide. The antiviral peptide is a peptide which can prevent or treat HIV, measles, RS V or Flaviviridae virus infection, and is referred to herein as a peptide inhibitor or a peptide of the present invention. "HIV" is a human immunodeficiency virus, which is a virus that causes immune deficiency by attacking CD4+ 10 cells in the body. The term "HIV" as used herein includes any HIV, including all populations and subtypes of HIV-1 and HIV-2 (branches). In several embodiments, the HIV is HIV-1. Measles, also known as rubeola, is a disease caused by a virus, especially caused by the paramyxovirus of the genus Morbillivims. Medication is transmitted through the respiratory (contact with the body fluids of the infected person's mouth and nose, direct contact or by droplets), and is highly contagious. 9〇% of people who are not immune will be infected with measles when they live together with the infected person. The respiratory syncytial virus (R S V) is a spherical or polymorphic enveloped virus (1〇〇_35〇N) with a single negative-vector linear RN A. There are two groups of people 2〇 RSV species, namely group A and group B. The majority of the A group is dominant. The present invention is directed to the treatment and prevention of two strains of RSV. RSV causes cold symptoms in adults and older children. However, young children may cause serious problems, including pneumonia and severe breathing problems. In rare cases, it can lead to death. According to the present invention, various types of HIV can be deactivated, or multiple types of infections can be treated and/or prevented because the evidence provided herein indicates that the peptide destroys or dissolves the virus without harming the mammal. cell. Thus, the procedure for inhibition of HIV infection by the peptide is not particularly phylogenetic, branch-specific or species-specific. Thus, the peptide has activity against a variety of HIV 5 types. Thus, one aspect of the invention relates to the treatment and prevention of infection by any of the HIV branch lines, race types, subtypes or germ lines. The invention is also directed to the deactivation of any branching, seed, subtype or germline HIV. Guidance for different branching lines can be found in the HIV Sequence Handbook 2002, edited by Kuiken C, Foley B, Freed 10 E, Hahn B, Marx P, McCutchan F, Mellors J, Wolinsky S, and Korber B, Los Alamos National Experiment Published by the Department of Theoretical Biology and Biophysics, LA-UR number 03-3564, herein incorporated by reference. Specifically, the HIV-1 branching lines A, B, C, and D and the outliers of multiple isolated strains can be found on pages 490 to 550; HIV-2 branches are A, 15 B, C and D and For the sequence data of multiple isolated plants, refer to pages Μ* to 578. The Hiv which can be activated, treated or inhibited by the present composition or method is usually HIV_1 or HIV-2, preferably HIV-. The HIV which can be deactivated, treated or inhibited according to the present invention is usually branched A, B, C or D. However, the composition of the present invention can also be used for the treatment or prevention of HIV-1 Μ or Ο group subtype or sub-type subtype infection. Members of the scorpion group are divided into nine equidistant phylogenetic subtypes. They are labeled Ai, A2, B, C, D, F1, F2, G, H, J and Κ, respectively. Sequence comparisons within any subtype or subtype are shared with other subtype sequences in the gene to share more genes. 56 200837075 The Flaviviridae is a spherical enveloped virus with a linear single-strand positive ^^ genomic body. The Flaviviridae includes the genus Flavivirus, Hepatitis, and Pesvirus. The present invention encompasses the treatment of Flaviviridae infections, including infections caused by any virus derived from any of the genus Flavivirus, Hepatitis and Pesvirus, and infections of viruses not designated in the Flaviviridae family. . For example, the peptide can be used to treat infections caused by the following viruvirus: sputum media encephalitis, Central European encephalitis, Far Eastern encephalitis, Rio Bravo, Sakamoto encephalitis, Kunjin (Kunjin) ), MuiTay Valley encephalitis, St. Louis encephalitis, West Nile encephalitis, Tyulenly, Ntaya, 10 UGanda S, dengue type 1, dengue fever Type 2, dengue type 3, dengue type 4, Modoc, and yellow fever. However, the peptide can be used to treat infections caused by the following prion viruses: bovine viral diarrhea virus type 1, bovine viral diarrhea virus type 2, porcine cholera (typical porcine fever virus) and Border disease (Border disease). )virus. In addition, the peptide can be used to treat infections caused by the hepatitis C virus classified as a hepatitis virus. The viruses of the non-designated genus of the Flaviviridae which can also be treated with the peptide of the present invention include: GB-A, GB-B and GB-C. To determine antiviral activity, peptides active against specific types of HIV, measles, RSV or Flaviviridae members, as well as appropriate 20 doses of such peptides, methods known in the art, including but not limited to, may be used. The dosage and method described. Viral infections in the presence or absence of a peptide of the invention can be assessed, for example, by measuring intracellular viral RNA levels, using the antiviral protein antibodies described herein, and detecting the number of viral proteins or viral lesions by immunoassay. Virus deactivation can be assessed using such methods as described herein. Resistance to peptides 57 200837075 Viral activity can also be determined using the vesicle release assay as exemplified herein. The peptide has antiviral activity if it can prevent or reduce viral infection or deactivate the virus for, for example, about 2 times or more than 2 times. For example, the peptide of the present invention can prevent or reduce viral infection by 2-5 fold, 5-10 fold or more than 10 fold. As exemplified below, the various peptides listed in Table 3 can inhibit viral infection by more than 10 fold, including, for example, having 8 called ID NO: 6, 8, 12, 13, 14, 24, 27, 30, 32 '43, 44 , 47, 48 and 53 peptides. The other peptides listed in Table 3 can inhibit viral infection by a factor of 5 to 1 fold, including peptides having SEq 10 ID NO: 21, 23, 28 and 37. The remaining peptides inhibit viral infection by at least 2-fold, while several remaining peptides inhibit viral infection by up to about 5-fold. These peptides have such viral infection inhibitory effects at concentrations of nanomolar and low micromolar concentrations. The peptide of the present invention is an amino acid polymer which is bonded by a guanamine linkage between an α-amine group and an α-carboxy group. Thus, the term "amino acid" as used herein refers to an alpha-amino acid. The amino acid included in the peptide of the present invention may be an amino acid or a D-amino acid. Further, the amino acid used in the peptide of the present invention may be a natural amino acid and an unnatural amino acid. Thus, the peptide of the present invention can be produced by genetically encoding an amino acid, a natural non-genetically encoded amino acid, or a synthetic amino acid. Examples of the amino acid representations used herein for the 20 genetically encoded L-amino acids and several uncoded amino acids are provided in Table 1. 58 200837075 Table 1

Amino acid single-letter symbols commonly used acronyms alanine A Ala arginine R Arg aspartate N Asn aspartic acid D Asp cysteine C Cys glutamine Q Gin glutamic acid E Glu glycine G Gly histamine Acid Η Hisisoleucine I lie leucine L Leu cleavage K Lys methionate Met phenylalanine F Phe bismuth citrate Pro leucine S Ser sulphate Thr tryptophan w Trp Tyrosin tyrosine Tyr proline V Val A-alanine Bala 2,3-diaminopropionic acid Dpr A-aminoisobutyric acid Aib N-mercaptoglycine (creatinine) MeGly ornithine Om Citrulline Cit 59 200837075 Amino acid single letter symbol commonly used abbreviation tert-butylalanine t-BuA third butyl glycine t-BuG N-methylisoleucine Melle phenylglycine Phg cyclohexyl Alanine Cha-Pro-Acetine Me Naphthyl-Alanine Nal 0-Bite Alanine 3-Benzothiophenylalanine 4-Chlorophenylalanine Phe(4-Cl) 2-Fluorophenylalanine Phe(2 -F) 3-fluorophenylalanine Phe(3-F) 4-fluorophenylalanine Phe(4-F) Penicillamine Pen 1,2,3,4-tetrahydro-isoindoline-3-carboxylate Acid T Ic A-2-mercaptoalanine Thi methionine hydrazine MSO high arginine Harg N-acetamido lysine AcLys 2,4-diaminobutyric acid Dbu N-aminophenyl phenylalanine Phe (pNH2) N-methylproline acid MeVal homocysteine Hcys homoserine Hser α-aminohexanoic acid Aha α-amino valeric acid Ava 2,3-diaminobutyric acid Dab 60 200837075 The peptide comprises at least 8 to about 50 amino acid residues, typically from about 14 to 40 amino acids, more typically less than about 35 amino acids or less than about 25 amino acids. The peptide of the present invention will be as small as possible while still maintaining the activity of substantially all of the larger peptides. Thus, the peptide of the present invention may be 8, 9, 10, 11, 12 or 13 amino acids. In addition, the length of the peptide selected by the skilled artisan is related to the stability and/or sequence of the peptide. Thus, for example, although peptide 1 (SEQ ID NO..43) has optimal antiviral activity when having about 18 amino acids, truncation by C terminal does not remove its anti-disease 10 toxic activity until five or less is deleted. Five amino acids. In spite of this, a peptide having a sequence different from SEQ ID NO: 43 has an optimum activity when it is longer than 18 amino acid or shorter than 13 amino acid. It may be due to the difference in sequence that results in the stabilization of the peptide or the modification of the secondary structure of the peptide. In addition, the peptide may be derived from an agent which enhances the stability or activity of the peptide. For example, the peptide can be modified to improve peptide activity and/or configuration stability via attachment of a dimethylaminonaphthalene moiety or via incorporation of a non-natural amino acid. The use of non-natural amino acids and dimethylaminonaphthalenesulfonyl moieties can also provide resistance to protease cleavage. In several cases, it is also desirable to combine two or more peptides into one peptide structure. 20 In addition, peptides from other HCV lines have excellent antiviral activity.

Including genotype IB (SWLRDVWDWICTVLTDFK, SEQ ID NO: 80); genotype 2A (SWLRDVWDWVCT1LTDFK, SEQ ID NO: 79); genotype 3A (DWLRII WDWVCSVVSDFK, SEQ ID 61 200837075 ID NO: 124); genotype 5A (TWLRAIWDWVCTALTDFK) , SEQ ID NO: 125); & *0S6A (SWLRDVWDWVCTVLSDFK, SEQ ID NO: 126) all have antiviral activity. The present invention is also directed to the antiviral peptide peptidomimetic agent of the present invention. 5 The structure of the phenopeptide action agent is similar to that of the peptide with a peptide bond, but one or more peptide bonds can be obtained by methods known in the art, such as _CH2NH-, one CH2S-, one CH2- CH2-------CH=CH—(Chuan and reverse), -COCH2-----CH(〇H)CH2-, and -CH2SO- are replaced by a bond. Such a 'pseudopeptide action agent is a peptide analog, such as the pharmaceutical industry commonly used as a 10 non-peptide drug drug' whose properties are similar to the properties of a peptide peptide (Fauchere, J.,

Adv. Drug Res" 15:29 (1986) and Evans et al, J. Med. Chem., 30:1229 (1987)). Advantages of Peptide Activator over Natural Peptide Examples include economical manufacturing High chemical stability, specificity change, and enhanced pharmacological properties such as half-life, absorbency, strength and efficacy. 15 In several embodiments, the amino acid residues of the peptide of the present invention are formed in solution. Amphipathic α _ helix structure. The term “α_helix” refers to the right-handed hover configuration. In a multi-peptide, the α-helical structure is obtained by hydrogen bonding between a main chain of an amino acid and an amino acid C0 group of an earlier 4 residues. The α-helix has 3.6 2 amino acid residues per enthalpy. Certain amino acid residues readily contribute to the formation of helical structures in the polypeptide, such as alanine, cysteine, leucine, methionine, face acid, glutamine, histidine, and lysine. ^ The formation of suspicion is also based on the solution, pH and temperature of the peptide. Thus, according to the present invention, the peptide of the present invention is an α-helix in an aqueous solution. 62 200837075 The aqueous solution is, for example, physiological pH & / or physiological salt. Typically, the amphiphilic alpha-helical structure of the peptide is detected at moderate temperatures, such as from about to about thieves, or from about room temperature to about body temperature. Thus, for example, the peptide has a 〇:-helix structure at physiological temperature and physiological pH. 5 α•Helical structures can be detected using methods known in the art, including but not limited to ® two-color spectroscopy (CD), nuclear magnetic resonance (NMR), crystal structure determination, and optical dispersion (ODD). As used herein, the term "amphiphilic" means that the alpha-helical peptide has a hydrophilic (or polar) surface and a water-repellent (or non-polar) surface, wherein such a "face" refers to the peptide. Longitudinal. When the α-helical peptide is viewed from the longitudinal axis (as shown in Fig. 6), the spiral wheel is conspicuous, and one side of the spiral wheel enclosing the longitudinal axis is composed of hydrophilic (or polar) residues, and The other side of the spiral wheel consists of water-repellent I* raw (or non-polar) residues. Thus, when the peptide of the present invention is located on the hydrophilic surface, the hydrophilic surface of the peptide tends to contact the hydrophilic surface. On the other hand, when 15 S faces the water repellent surface, the water repellent surface of the peptide of the present invention tends to contact the water repellent surface. In the amphipathic α-helical peptide, the hydrophilic surface and the water-repellent surface of the α-helix can be identified based on the nature of the amino acid present. The hydrophilic surface of the ^^ spiral will consist of more hydrophilic, charged and/or polar amine bismuth acids present on the water repellent surface. The water-repellent surface of the amphipathic 0^ helix is composed of a water-repellent amino acid and/or a non-polar amino acid which can assist in the insertion of the lipid bilayer. The water repellent surface has one or more hydrophilic amino acids or polar amino acids, as long as sufficient amounts of non-polar private acid are present to allow membrane insertion. Usually, most of the amino acid residues on the hydrophilic surface of the α-helix are charged amino acids or polar amino acids, and most of the amino acid residues on the water surface of the snail 63 200837075 are Non-polar amino acid. Thus, in various embodiments, the hydrophilic surface of the 〇:-helix is composed of a charged amino acid or a polar amino acid, and the water-repellent surface of the α-helix is composed of a non-polar amino acid residue. For example, reference to the helical wheel of peptide 1 (SEQ ID NO: 43), shown in Figure 6A. Whether a given peptide sequence has a sufficient number of non-polar amino acids to allow membrane insertion can be determined using methods well known in the art, including but not limited to the release of vesicle dyes as described in the examples herein. method. In addition, whether the peptide has an amphipathic α-helical structure can be determined using software available on the Internet 10, such as http://cti.itc.virginia.edu/~cmg/

Demo/wheel/wheelApp.html (last visit August 15, 2006) http://www.bioinf.man.ac.uk/~gibson/HelixDraw/hdixdraw-h tml (last visit August 15, 2006) day). A schematic representation of the amphipathic a-helical structure showing the peptide of SEQ ID NO: 43 is shown in Figure 6A. 15 For an example of the peptide of the present invention, refer to Table 3. Other peptides of the invention include comparing the peptides shown in Table 3 with conservative amino acid substitutions. The peptide of the present invention also includes such peptides having an amino acid composition similar to the peptide shown in Table 3. These peptides include the peptides having the sequences of SEQ ID NOS: 96, 97 and 98 and are shown in Table 11. These sequences correspond to the variants of SEQ ID NO: 43 that correspond to 20 different strains or their constituents of 8: 卩 still NO: 43 "disintegrated" variants. A retro- or mutant variant such as the peptide of SEQ ID NO: 43 has an amino acid composition similar to the original peptide (SEQ ID NO: 43), but the amino acid sequence is the sequence of the original peptide Upside down. The peptide, such as the transcribed variant of SEQ ID NO.43, also has an amino acid composition similar to that of the original Shengxin (seqid 64 200837075 NO: 43), but the sequence of the amino acid will be broken up or mixed without change. The relative position of the water-repellent residue and the hydrophilic residue. Thus, the peptide system belonging to the "water-repellent-dispersed" variant of SEQ ID NO: 43 has the same amino acid composition as SEQ ID NO: 43. However, the order of the water-repellent amino acid 5 residue will be changed without changing the relative position of the water-repellent residue and the hydrophilic residue in the sequence. Therefore, the amphipathic line of the mutant peptide is similar to the two parents of the original peptide. Sex. Similarly, the peptide of the "hydrophilic disintegrated" variant of SEQ ID NO: 43 has the same amino acid composition as SEQ ID NO: 43, but the order of the hydrophilic amino acid residues will be changed. The relative position of the water-repellent residue and the hydrophilic residue of the 10 parts in the sequence is not changed, so the amphipathic line of the mutant peptide is similar to the amphipathicity of the original peptide. Generally, in the case of hydrophilic (polar) amino acids, the words "break up" or "dissipated" are used to indicate that the position of each hydrophilic (polar) amino acid remains constant, and any other hydrophilicity (polarity) The amino acid can be located at this position. Similarly, in the case of water-repellent (non-polar) amine 15 acid, the words "breaking up" or "breaking" are used to indicate that the position of each water-repellent (non-polar) amino acid remains constant, any other. A water repellent (non-polar) amino acid can be located at this position. Thus, the peptide of the present invention has the same amino acid sequence as the sequence shown in Table 3 and a variant of these sequences. These variants may be derived from one or more amino acid truncations, conservative substitutions, only the relaxation of hydrophilic amino acids, the scattering of only water-repellent residues within a sequence, hydrophilic amino acids and Dispersion of both water-repellent amino acids, replacement of the native amino acid with an unnatural amino acid or other modification such as dimethylamine sulfonium sulfonation. These variant peptides are further described in the next section. 65 200837075 Peptide homologs and variants The present invention encompasses a wide variety of peptide homologs and variants. For example, although the antiviral peptide is originally isolated from the hepatitis C genotype la (H77), similar peptides from other HCV lines have excellent antiviral activity including: 5 genotype IB (SWLRDVWDWICTVLTDFK, SEQ ID NO: 80); genotype 2A (SWLRDVWDWVCTILTDFK, SEQ ID NO: 79); genotype 3A (DWLRII WDWVCSVVSDFK, SEQ ID NO: 10 123); genotype 4A (SWLWEVWDWVLHVLSDFK, SEQ ID NO: 124); genotype 5A ( TWLRAIWDWVCTALTDFK, SEQ ID NO: 125); and

15 Genotype 6A (SWLRDVWDWVCTVLSDFK, SEQ ID NO: 126) all have antiviral activity. Thus, the invention is also directed to a peptide homologue of the active peptide disclosed herein. The peptide homologue is the peptide sequence derived from the HCV isolate from the H77 isolate of SEQ ID NO. Thus, the peptide of the present invention may be 20 a peptide homolog having the amino acid sequence of any one of SEQ ID NO: 4-61. Thus, for example, a peptide homologue of the invention has a SEq still NO>62' which is a homologue of the peptide seq id NO:6. LYGNEGLGWAGWLLSPRG (SEQ ID NO: 62). The peptide inhibitor sequence SEQ ID NO: 62 is found in the HCV polyprotein sequence 66 200837075 column SEQ ID NO: 2 and 3. Another peptide inhibitor homolog of the invention has SEQ K) NO: 63 or 64 which is a homolog of the peptide SEQ ID NO: 8. IFLLALLSCITVPVSAAQ (SEQ ID NO: 63); 5 IFLLALLSCLTIPASAYE (SEQ ID NO: 64). The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another peptide inhibitor homolog of the invention has SEQ ID NO: 65 or 66 which is a homolog of the peptide SEQ ID NO: 12. 10 MSATFCSALYVGDLCGGV (SEQ ID NO: 65)

GAAALCSAMYVGDLCGSV (SEQ ID NO: 66) The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another peptide inhibitor homolog of the invention has SEQ ID NO: 15 67 or 68 which is a homolog of the peptide SEQ ID NO: 13. ALYVGDLCGGVMLAAQVF (SEQ ID NO: 67) AMYVGDLCGSVFLVAQLF (SEQ ID NO: 68) The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another homologous peptide inhibitor homolog of the invention has SEQ ID NO: 69 or 70 which is a homolog of the peptide SEQ ID NO: 14. IIDIVSGAHWGVMFGLAY (SEQ ID NO: 69) VVDMVAGAHWGVLAGLAY (SEQ ID NO: 70) 67 200837075 The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another peptide inhibitor homologue of the invention has SEQ ID NO: 71 or 72 which is a homolog of the peptide SEQ ID NO: 24. 5 VDVQYMYGLSPAITKYVV (SEQ ID NO: 71) YLYGIGSAVVSFAIKWEY (SEQ ID NO: 72) The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another peptide inhibitor homolog of the invention has SEQ ID NO: 10 73 or 74 which is a homolog of the peptide SEQ ID NO: 27. WMLILLGQAEAALEKLVV (SEQ ID NO: 73) WMMLLIAQAEAALENLVV (SEQ ID NO: 74) The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another homologous peptide inhibitor homolog of the invention has SEQ ID NO: 75 or 76 which is a homolog of the peptide SEQ ID NO: 30. GVVFDITKWLLALLGPAY (SEQ ID NO: 75); ELIFTITKILLAILGPLM (SEQ ID NO: 76). The sequences of such peptide inhibitors are found in the HCV polyprotein sequence SEQ ID 20 NO: 2 and 3. Another peptide inhibitor homolog of the invention has SEQ ID NO: 77 or 78, which is the peptide SEQ ID NO: 32 homologues. VSQSFLGTTISGVLWTVY (SEQ ID NO: 77); ATQSFLATCVNGVCWTVY (SEQ ID NO: 78). 68 200837075 The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another peptide inhibitor homolog of the invention has SEQ ID NO: 79 or 80 which is a homolog of the peptide SEQ ID NO: 43. 5 SWLRDVWDWVCTILTDFK (SEQ ID NO: 79); SWLRDVWDWICTVLTDFK (SEQ ID NO: 80). The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO.. 2 and 3. Another peptide inhibitor homologue of the invention has SEQ ID NO 10180 or 82 which is a homolog of the peptide SEQ ID NO: 44. DWVCTIUrDFKNWLTSKL (SEQ ID NO: 81); DWICTVLTDFKTWLQSKL (SEQ ID NO: 82). The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. Another homologous peptide inhibitor homolog of the invention has SEQ ID NO: 83 or 84 which is a homolog of the peptide SEQ ID NO: 47. ASEDVYCCSMSYTWT (SEQ ID NO: 83); EDDTTVCCSMSYSW (SEQ ID NO: 84). The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID 20 NO: 2 and 3. Another peptide inhibitor homologue of the invention has SEQ ID NO: 85 or 86 which is a homolog of the peptide SEQ ID NO..53. 69 200837075 CTMLVCGDDLVVICESAG (SEQ ID NO:85); PTMLVCG DDLVVISESQG (SEQ ID NO:86) ° The sequences of these peptide inhibitors are found in the HCV polyprotein sequences SEQ ID NO: 2 and 3. The 5 peptide mutant is any peptide having a different amino acid sequence from the amino acid sequence of the HCV isolate. Thus, the peptide of the present invention has a mutant sequence obtained by substituting a conservative amino acid. Amino acids which can be substituted with each other generally belong to a similar class or subclass. As known to those skilled in the art, amino acids can be placed in different classes depending on the chemical nature and physical properties of the amino acid branch. For example, several amino acids are generally considered to be hydrophilic or polar amino acids, while other amino acids are considered to be water-repellent or non-polar amino acids. The polar amino acid includes an acid group having an acidic, basic or hydrophilic branch, and the non-polar amino acid includes an amino acid having an aromatic branch or a water-retained branch. The non-polar amino acid can be further subdivided to include an aliphatic amino acid 15 and the like. The amino acid class as used herein is defined as follows. "Non-polar amino acid" means having a branch and physiology? 11 is uncharged, it is non-polar, and the amino acid normally rejected by aqueous solutions. Examples of genetically encoded water-repellent amino acids include Ala, lie, Leu, Met, Trp, Tyr, and Val. Examples of non-genetically encoded non-polar amino acids include 6; 311 VIII, cha and Nle. 20 "Aromatic amino acid" means a non-polar amino acid having a branched chain containing at least one ring (aromatic group) of a 5-electron system. The aromatic group may further be a substituent such as an alkyl group or an alkene. Substituted by alkynyl, alkynyl, hydroxy, sulfonyl, nitro, and amine groups, etc. Examples of genetically encoded aromatic amino acids include phenylalanine, tyrosine, and tryptophan. Common non-genetically encoded aromatic amines Acid acid package 70 200837075 includes phenylglycine, 2-naphthylalanine, a-2-nonyl alanine, 〗 2,3,4_ four wind-iso-na-3 acid, 4-chlorobenzene Alkalamine, 2-chlorophenylalanine, 3-chlorophenylalanine, and 4-chlorophenylalanine. "Amino amino acid" means a saturated or unsaturated linear, branched 5^ ring. Hydrocarbon branched non-polar amino acids. Examples of aliphatic amino acids include Ala, Leu, Val and lie. Examples of non-coding aliphatic amino acids include Nle. '"Polar amino acid" means a branched hydrophilic amino acid which is charged or uncharged at physiological pH and which has a bond, which is shared by two 10 atoms - It is maintained more closely by one of the atoms. The polar amino acid is usually hydrophilic, indicating that one branch is an amino acid that is attracted to the aqueous solution. Examples of genetically encoded polar amino acids include the day. Winter oxime, cysteine, glutamine, lysine, and serine. Examples of non-genetically encoded polar amino acids include citrulline, homocysteine, N-acetamide, 15 amino acid and "Acidamine" refers to a hydrophilic amino acid having a branched pK value of less than 7. Acidic amino acids typically have a negatively charged branch due to the loss of hydrogen atoms. . Examples of genetically encoded acidic amino acids include aspartic acid (aspartate) and glutamic acid (glutamate). 20 "Indicative amino acid" means a hydrophilic amino acid having a branched chain pK value of more than 7. Since the basic amino acid is combined with a hydrogen ion, it has a positively charged branch in the physiological state. Examples of genetically encoded basic amino acids include arginine, lysine, and histidine. Examples of the non-genetically encoded basic amino acid include amino acid ornithine, 2,3-diaminopropionic acid, 2, towedibutyric acid, and high-precision 71 200837075 aminic acid. "Free amino acid" means an amino acid which is physiologically pfi-chargeable. Such free amino acids include acidic amino acids and in situ amino acids, such as hydrazine-aspartic acid, D- faceamine, D-histamine, D-arginine, lysine, D- Hydroxy 5-amino acid, D-ornithine, L-aspartic acid, L- face acid, L-histamine, L-arginine, L-lysine, L-hydroxy lysine or L-ornithine. As the skilled artisans understand, the aforementioned classification is not absolute. Several amino acids have more than one characteristic property and can therefore be included in more than one class. For example, tyrosine has a non-aromatic aromatic ring and a polar hydroxyl group. Thus, tyrosine 10 has several properties that can be described as non-polar, aromatic, and polar. However, the non-polar ring is the main control. Similarly, cysteine has non-polar properties in addition to its formation of disulfide linkages. Although not strictly classified as a water-repellent amino acid or a non-polar amino acid, cysteine can be used to provide water-repellent or non-polarity to the peptide in many cases. 15 The classification of the aforementioned genetically encoded amino acids and non-coding amino acids is summarized in Table 2 below. However, it should be understood that Table 2 is for illustrative purposes only and is not an exclusive form that may include the amino acid residues of the peptides and peptide analogs described herein. Other amino acid residues useful in the manufacture of the peptides described herein can be found, for example, in Fasman, 1989, CRC Handbook of Applied Biochemistry and Molecular Biology, CRC Publications, and references cited therein. Another source of amino acid residues is provided by the RSP amino acid analog company website. 〜1^111丨110-3(^(18.〇0111). Amino acids not specifically mentioned here. It is conveniently classified into the aforementioned categories based on known performance and/or comparison of the characteristic chemical and/or physical properties of the amino acids specifically identified herein. 72 200837075 Table 2 Classification Genetic coding Non-genetically encoded non-polar aromatic F, Y, W Phg, Nal, Thi, Tic, Phe (4-Cl), Phe (2-F), Phe (3-F), Phe (4-F), pyridyl Ala, benzophene Ala aliphatic AV, L, I t-BuA, t-BuQ Melle, Me, MeVal, Cha? bAla, MeGly, Aib Other non-polar H QP polarity ~ Acid D, E testability ------- h, k,r Dpr,Om,hArg,Phe(p-NH2), DBU, A2BU Neutral polar cysteine---- H; Y,Q,N,D,E,H, JK,C__ Cit , AcLys, MSO, hSer, Om, Hcys C Pen, hCys, β-methyl Cys. In several embodiments, the hydrophilic or polar amino acids encompassed by the present invention include, for example, arginine, aspartame, aspartame Acid, cysteine, glutamic acid, Chuamine, histidine High cysteine, lysine, hydroxy lysine, ornithine, serine, threonine, and structurally related amino acids. In a real case, 'polar amino acids are free Amino acids such as arginine, aspartic acid, sulphate, histidine, hydroxy lysine, lysine or ornithine. Examples of useful water-repellent or non-polar amino acid residues include, for example, C. 1 valine, valine, leucine, methionine, isoleucine, phenylalanine, tryptophan, tyrosine, etc. In addition, the amino acid sequence of the peptide can be modified, Thus, a peptide variant is obtained which comprises at least one amino acid residue in the peptide in place of another amino acid residue, including substitution using the D form rather than the L form. 73 200837075 One or more may be exchanged for the other to alter, enhance or retain the biological activity of the peptide. Such variants, for example, have at least about 10% of the biological activity of the corresponding non-mutant peptide. Conservative amino acid substitution, ie amino acid 5 substitution with similar chemical and physical properties, as previously mentioned Thus, for example, a conservative amino acid substitution involves aspartic acid exchange glutamic acid; lysine exchange arginine or histidine; a non-polar amino acid (alanine, isoleucine, leucine, Methionine, phenylalanine, serine, tyrosine, proline) exchange for another non-polar amino acid and a polar amino acid (day 10, aspartame, face acid, Face amine, glycine, serine, threonine, etc. are exchanged for another polar amino acid. When a substitution is introduced, the variant can be tested to verify or determine its biological activity. For example, in several embodiments, the peptide of the present invention may comprise any of the sequences of Formula IX-XIII: 师〇;: XMirXtoii^Cm^^Cia^XMir (SEQ © MO: 113) 114)

Xaair^i4-XM|f ^BQ ID NOi 1 IS)

Km\icr Xmj i - Kill 15 (SEQ ID NO: 116) where:

Xaai ' Xaa4 ^ Xaa5 ^ Xaa8 Λ Xaan ' Xaa^ Λ Xaai5 ^ Xaai6 and Xaa18 are each a polar amino acid; and 74 200837075

Xaa2, Xaa3, Xaa6, Xaa7, Xaa9, Xaa10, Xaa13, Xaa14,

Each of Xaa17 is a non-polar amino acid, respectively. In other embodiments, the peptide may have additional peptides and sequences at the N-terminus or C-terminus. Thus, for example, the present invention provides a fusion peptide formed by attaching a μ-amino acid peptide (N-terminal peptide) 5 to the N-terminus of any of the peptides of Formula IX to ΧΙΟ. The 14 amino acid N-terminal peptide has the structural formula: Rx-Ry-Ry-Rx-Ry-Ry-Rx.Rx.Ry.Ry-Rx.Rx.Ry.Rx (SEQ ID NO: 117), wherein each RX is a polar amino acid, and each of them is a non-polar amino acid. The present invention also provides a fusion peptide formed by attaching a 12 amino acid peptide (C-terminal peptide) to the C-terminus of the formula XIII peptide. The resulting fusion peptide has the structural formula of formula XIV:

Xaai2'XHEi3_X (10) 14*" Χ3Ει5~Χηηι6-Χ (10) 17-X(10) is -X(10) 19-X(10)2〇_Xaa2i-Xaa22-15 Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa3〇(SEQ ID NO : 118), XIV where:

Xaa!, Xaa4, Xaa5, Xaa8, Xaau, Xaa12, Xaa15, Xaa16, Xaa18, Xaa19, Xaa22, Xaa23, Xaa26, Xaa29, and Xaa30 are each a polar amino acid; and 20 Xa&2, Xaa3, Xaa6, Xaa7 , Xaa9, Xaa10, Xaa13, Xaa14,

Xaa17, Xaa20, Xaa2i, Xaa24, Xaa25, Xaa27, and Xaa28 are each a non-polar amino acid. The present invention also provides a fusion peptide having a sequence corresponding to one of the amino acid N-terminal peptides of SEQ ID NO: 117, which is linked to one of the N-terminal peptides of the formula XIV peptide, 75/0737075. In another embodiment, the peptide of the present invention is a peptide comprising at least 14 contiguous amino acids of any of the foregoing peptides. The peptide variant can also be obtained from a sequence of internal hydrophilic residues and / or 5 water-repellent residues "break up", as long as the amphiphilic α-helical secondary structure of the peptide in solution can be maintained. . Method of Producing the Peptide of the Invention In the context of the present invention, the "isolated" peptide is a peptide which exists in addition to its natural environment and is therefore not a natural product. The isolated peptide may be present in purified form or may be present in a non-native environment, such as in the presence or absence of a cell, or other active ingredient or inactive ingredient may be present in the composition in the composition. In one embodiment, the "isolated" peptide does not contain the peptide from which the peptide was originally derived, and is naturally at least partially sequenced from the peptide (ie, at the top of the peptide) And the sequence of the C-end). The "purified 15" peptide is substantially free of other cellular material or culture medium (when manufactured by the heavyweight technique), or substantially free of chemical precursors or other chemicals (when chemically synthesized). Thus, the purified peptide preparation is at least Na, to ^ 60 /. ' to / 7G/. 'At least 8 ()%, or at least 9q% by weight of the skin. Purity can be determined using methods known in the art, including but not limited to methods using chromatography or polyacrylamide gel electrophoresis. The peptide or a variant thereof can be synthesized in a test tube, for example, by a solid phase peptide method, or by a recombinant (10) eight technique. The solid phase peptide synthesis method is an established and widely used method, and the method is described in the following references: et al., solid phase wins skin 76 200837075 Synthesis 'WH Freeman Co., San Francisco ( 1969); Merrifield, I. Am. Chem. Soc. 85 2149 (1963); Meienhofer, "The Installation of Hormone Proteins and Peptides"; CH·Li, Vol. 2 (Academic Press, 1973), pp. 48-267 And Bavaay and Merrifield, "Split Peptides", ed. 5 E. Gross and F. Meienhofer, vol. 2 (Academic Press, 1980), pp. 3-285. These peptides can be further subjected to immunoaffinity chromatography or ion exchange column sorting; ethanol precipitation; reverse phase HPLC; chromatography on cerium oxide or anion exchange resin such as DEAE; chromatography; SDS-PAGE; Ammonium precipitation; gel filtration using, for example, Sephadex G-75; 10 ligand affinity chromatography; or purification by crystallization or precipitation from a non-polar solvent or a mixture of non-polar/polar solvents. It is preferred to purify by crystallization or precipitation. The peptide of the present invention may be a cyclic peptide as long as it retains antiviral activity. Such cyclic peptides are typically produced by linear peptides by covalent attachment to the terminal carboxylate via the amine terminus. To ensure that only the terminals are joined, the amine branch 15 chain and the carboxylic acid branch can be protected using commercially available protecting groups. In several embodiments, one skilled in the art can choose to cyclize a peptide branch to one of the amine or carboxy terminus, or to another amino acid branch. In this case, the protecting group can be used again to guide the cyclization reaction as needed. The victory of the moon can be done using existing procedures. For example, benzobis 20 azole-1 thio-pyrrolidinyl hexafluorophosphate scale (PyB〇p, Novabiochem) (5 equivalents relative to the crude peptide) and N can be used. A mixture of N-diisopropylethylamine (DIEA, Fisher) (40 equivalents) was cyclized at dimethyl cyanoamine at a peptide concentration of 1-5 mM. The number of diea has been adjusted to achieve a nominal pH of 9-10. The reaction can be followed by any convenient means such as 77 200837075 by MALDI-MS and/or HPLC. The N-coradyl derivative of the amino group of the peptide or peptide variant can utilize N-mercapto protected amino acid for final condensation, or via deuterated protected peptide or unprotected peptide preparation. The indole-indenyl derivative can be prepared, for example, by deuteration of the free form 5 via a base peptide or a peptide resin. The deuteration can be carried out using a standard acidifying agent such as a mercapto halide, an anhydride, a mercapto-based yttrium or the like. If desired, both Ν-醯 and 0-醯 can be performed together. The salt of the carboxyl group of the peptide or peptide variant of the present invention can be prepared in an ordinary manner by contacting the peptide with one or more equivalents of a desired base, such as a metal hydroxide such as sodium hydroxide; Salt test or bicarbonate test such as sodium sulphate or sodium sulphate; or amine test such as triethylamine, triethanolamine and the like. An acid addition salt of a peptide or a variant peptide, or an acid addition salt of an amino group residue of a peptide or a variant peptide, may be via a peptide or an amine with one or more equivalents of a mineral acid or organic It is prepared by contacting an acid such as hydrochloric acid. The ester of the carboxyl group of the peptide can be prepared by any of the usual methods known to the art. Method of Use The peptide of the present invention can be used to prevent, treat or ameliorate any viral infection by any of the human immunodeficiency virus (HIV), aphrodisiac or respiratory fusion virus, and from the 20 families of flaviviruses. Furthermore, the peptide of the present invention can be used to activate any of these viruses in vivo, in vitro or in vitro. The peptide of the present invention can be used for preventing, treating or ameliorating infection by any HIV, aphrodisiac virus or RSV virus and any Flaviviridae virus; and can prevent, treat or ameliorate related diseases. In this way, the peptide can be used as a therapeutic agent to prevent sexually transmitted HIV, to treat and inhibit HIV disease, to inhibit the reproduction of HIV, to reduce the load of HIV, and to enhance the CD4+ response against HIV. Enhance the CD8+ response against HIv to cure HIV infection, improve the safety of blood and blood products used in blood transfusions, and increase the safety of bed test samples. The skin of the present invention can also be used to deactivate virus in body fluid samples and tissue samples, and to inhibit or prevent infection with those who must handle such samples. Similarly, the peptide of the present invention can be used to prevent, treat or ameliorate infection by Flaviviridae viruses, which include the Flavivirus, Pestivirus 10 genera, and Hepatitis virus as previously described. Members of the Flavivirus include sputum media encephalitis, CEIBS, Far Eastern encephalitis, Leo Bravo, Japanese encephalitis, Kunjing, Murray Valley encephalitis, St. Louis encephalitis, West Nile encephalitis, and sudden Larry, Taya, Uganda S, Dengue Type 1, Dengue Type 2, Dengue Type 3, Dengue Type 4, Modoc, and Yellow Fever. The prion belongs to 15 members including bovine viral diarrhea virus type 1, bovine viral diarrhea virus type 2, swine disorder (typically hot virus) and Bord's disease virus. Hepatitis virus genus includes hepatitis C virus. Additional members of the Flaviviridae include unspecified GB-A, GB, and GB-C. Members of the Flaviviridae virus are known to cause a variety of diseases, including, for example, dengue fever, hepatitis C, Japanese encephalitis, Kyasan^ forest disease, Murray Valley encephalitis, St. Louis encephalitis, and sputum encephalitis. , West Nile encephalitis and yellow fever. The peptide of the present invention can be used to prevent, treat or ameliorate infection by members of the Flaviviridae virus and related disease states. Thus, various application examples of the present invention include, but are not limited to, use as dengue fever, dengue fever, dengue 79 200837075 heat shock syndrome, Japanese encephalitis, Jiasha Nuersen eucalyptus, Mu Ruigu encephalitis, A therapeutic agent for patients with St. Louis encephalitis, meningococcal encephalitis, and chronic hepatitis C infection; prevention of infection of the graft during liver transplantation, prevention of miscellaneous, and increased safety of blood and blood products for blood transfusion Sex, as well as the safety of the 5 high clinical test samples. In one embodiment, the invention provides a method for preventing, inhibiting or ameliorating a viral infection of a mammalian cell, such as a human cell; or preventing, inhibiting, treating or ameliorating a mammal, such as a human immunodeficiency virus, an anesthetic A method of acute infection or slow 10 infection of a virus or a flavivirus. As used herein, the term "prevention" is intended to include administration of a peptide of the present invention to a milk-removing animal, such as a human, which has been exposed to or has been exposed to a human immunodeficiency virus, for use in suppressing infection. Mammals that may be exposed to HIV include, but are not limited to, those living in areas where the virus is already prevalent or common in the virus, such as Africa, Southeast Asia, China, South Asia, Australia, India,

US, Russia, Muse, and Central and South American countries. Feeding animals that may be exposed to HIV also include recipients who have been donated to body tissues or body fluids, such as blood or blood; or a variety of components such as blood stasis, platelets, or stem cells; Tissue and body fluid medical, clinical 20 and dental staff. Mammals that have been exposed to HIV include, but are not limited to, those who are exposed to infected individuals or those who have been exposed to HIV, such as blood. Furthermore, prophylaxis is intended to include the administration of a peptide of the present invention to a mammal, such as a human, that may be exposed to or has been exposed to members of the Flaviviridae family, to inhibit the infection of the Flaviviridae virus in 200837075. Mammals that may be exposed to the Flaviviridae virus include, but are not limited to, mammals living in sick or common areas such as the tropics, Southeast Asia and the Far East, South Asia, Australia and Pakistan, Asia and Guinea, and the United States. , Russia, Germany, and South America. Mammals that may be violent to the Flaviviridae virus include those who have been bitten by deer or forest monuments or mosquitoes; recipients of body fluids, such as blood 'night' or one or more blood components such as plasma, Platelet, or stem cell donor; and medical or dental practitioners who handle body tissues and body fluids, including mammals who are secretive to flaviviruses, including but not limited to those who have been exposed to sputum or HCV or any other flavivirus A body tissue or body fluid of a virus, such as a mammal of blood. Further, "prevention" is intended to include administration of a peptide of the present invention to a Wei animal, such as a human, that may be exposed to or has been exposed to a respiratory virus, such as a human, to suppress measles virus or RSV infection. Treating or treating HIV infection, aphrodisiac infection, lining infection, or yellow disease virus lines: t® includes alleviating the load of Hess, or improving or alleviating at least one symptom of the typical and infectious phase. This treatment also includes improving or reducing more than one symptom. It sounds like treating _ faithfulness to suppress viral infections and/or eliminate the symptoms associated with infection. Symptoms or characterization of viral exposure or viral infection are characterized by specific symptoms or characterization of the particular infection and are known to those skilled in the art. Early silk infections include flu-like symptoms that are exposed to three to six weeks of disease reduction. This disease is referred to as acute mv syndrome, including fever, headache, tiredness L, abdominal filling, and lymphadenopathy. These 81 200837075 Symptoms usually disappear within weeks to weeks and are often misdiagnosed as other viral infections. During this acute phase, the amount of virus in the body is high and the virus spreads to the body. The p-knife, especially the lymphoid tissue. After this stage, the infected person is more likely to spread the infection to others. Then along with the body's immune system Coordinating operations at the beginning of 5, the amount of virus is reduced. Adults who have entered the body for the first time or even a decade or more, or congenitally infected with a virus in two years, more persistent or more serious symptoms may not It will float on the face. This "asymptomatic" infection period varies from person to person. Some people start to have symptoms within a few months, others may still have no symptoms for more than a decade. However, during the “no symptoms” period, the virus actively breeds, infects and kills cells of the immune system. Symptoms associated with infections caused by members of the Flaviviridae family, such as dengue or hemorrhagic dengue, are caused by one of four flavivirus subtypes. Symptoms of these diseases include sudden onset of fever 'severe headache, joint and muscle pain and rash, as well as high fever, thrombocytopenia and blood concentration. The 15-bed instructions also include high fever, bleeding spots with thrombocytopenia and reduced white blood cells, and bleeding tendency. Japanese symptoms of brain inflammation include fever, headache, neck stiffness, cachexia, hemiplegia, convulsions, and elevated body temperature. Japanese encephalitis can be diagnosed by detecting antibodies in serum and cerebrospinal fluid. Symptoms of Jiashaur's forest disease include high fever, headache, bleeding from the nose and throat, and vomiting. Sympathy for St. Louis 20 includes fever, headache, stiff neck, stupor, loss of orientation, fainting, tremors, occasional convulsions, and spastic paralysis. Symptoms of Murray's encephalitis include fever, seizures, nausea, and diarrhea in children; and headaches, lethargy, and confusion in adults. West Nile virus infections include flu-like symptoms, malaise, fever, loss of appetite, nausea, vomiting, eye pain, headache, myalgia, rash 82 200837075 and lymphadenopathy, as well as encephalitis (inflammation of the brain) and meningitis (brain And the lining of the spinal cord is inflamed), pseudocerebroenitis, temporary insomnia, seizures and coma. West Nile infection can be detected by ELISA for detection of antibodies in blood and cerebrospinal fluid. Yellow fever symptoms include fever, myalgia, headache, back pain, 5 red tongue, facial flushing, red eye, gastrointestinal bleeding, bloodshot throat, jaundice, liver failure, renal insufficiency with proteinuria, blood pressure Low, dehydration, spasms, seizures and coma. Symptoms of hepatitis C infection include, but are not limited to, liver infection, loss of appetite, burnout, abdominal pain, jaundice, flu-like symptoms, itching, myalgia, joint pain, intermittent low fever, sleep disturbance, nausea, 10 indigestion, cognition Change, depression, headache and emotional changes. HCV infection can also be diagnosed by detecting viral antibodies and detecting liver inflammation, cirrhosis, portal hypertension, thyroiditis, cryoglobulinemia, and renal glomerulonephritis by biopsy. In addition, HCV infection can be diagnosed. In addition, abnormal liver enzymes or liver function tests based on medical history, routine blood tests can be used to detect exposure to viruses or 15 infected viruses or to identify individuals at risk of exposure to HCV. Usually, infection by a member of the Flaviviridae can be diagnosed using an ELISA for detecting a viral antigen or an antiviral antibody, an immunological glory for detecting a disease antigen, and a polymerase chain reaction (PCR) for detecting a viral nucleic acid. Measles symptoms include fever for at least three days and cough, runny nose, and conjunctival inflammation. Fever up to 40 ° C (l 〇 4 ° F). The presence of Koplik's s spots in the mouth is a special symptom of the virus (diagnostic symptoms), but it does not occur frequently. Even true measles cases do not necessarily exist frequently because of the Copril point. It is temporary and disappears within a few days after its appearance. A characteristic rash of measles is typically described as a comprehensive papular-like red that begins several days after the onset of fever 83 200837075 A plaque rash. It begins to appear on the head and then spreads over most of the body surface, often causing itching. The treatment is called "freckle", and the color changes from red to dark brown before disappearing. RSV causes respiratory infections in patients of all ages. Rsv is the main cause of lower respiratory tract infections in infants and children. Symptoms of RSV include recurrent wheezing, and severe RSV infection in infants of the first few months of age may cause asthma. Methods of preventing, treating or ameliorating acute viral infections or chronic viral infections include contacting the cells with an effective amount of a peptide of the invention, or administering a therapeutically effective amount of a peptide of the invention to a mammal, such as a human. The deactivation of the diseased bone comprises contacting the virus with an effective amount of the peptide of the present invention, which can be administered in a variety of ways. The route of administration includes but not

The peptide of the present invention is swallowed by 摅 ϋ,, -

20 84 200837075 Several factors that determine the appropriate dosage are well known to those skilled in the art and can be solved by routine experimentation. For example, the determination of physicochemical properties, toxicological properties, and pharmacodynamic properties can be achieved using standard chemical assays and bioassay analyses well known in the chemical, pharmacological, and toxicological communities and through the use of mathematical modeling techniques. Therapeutic appliances and medications are extrapolated from the results of this specialized technique and extrapolated through the use of appropriate pharmacokinetic and/or pharmacodynamic models. Other factors will be determined based on the parameters of the individual patient, including age, physical condition, size, weight, condition of the treatment, severity of the condition, and any concurrent treatment. The dosage is also determined by the skin to be used, the purpose of the treatment or prevention, and whether the skin has been chemically modified. These factors are conveniently determined by the clinician using a viral infection model such as the Hey Cell Culture/JFH_1 infection model described herein or other viral models or test systems readily available in the artisan community. 15 The exact dose of the patient will be determined by the clinician. However, in order to achieve the desired effect, the peptide of the present invention, a variant thereof, or a combination thereof may be administered in a single dose or in divided doses, for example, at least about 1 mg/kg to about 500 to 750 mg/kg, At least about 0. 01 mg/kg to about 300 to 500 g/kg, at least about 1 mg/kg to about 100 to 300 mg/kg, 20 or at least about 1 mg/kg to about 50 to 100 mg/ Kilograms of body weight, but other doses can achieve beneficial effects. The absolute weight of a given peptide contained in a unit dose can vary widely. For example, from about 0. 01 grams to about 2 grams or from about 0.1 milligrams to about 500 milligrams of at least one peptide of the present invention or a plurality of specific peptides for a particular cell type. In addition, the unit dosage can be varied from about 0.10 grams to about 50 grams, from about 1 gram to about 35 grams, from about 1 gram to about 25 grams, from about 5 grams to about 12 grams. From about 5 grams to about 8 grams, from about 5 grams to about 4 grams, or from about 5 grams to about 2 grams. The dose per peptide of the peptide of the invention may also vary. Such doses per dose 5 may range from about 0.1 g/day to about 50 g/day, from about 01 g/day to about 25 g/day, from about 〇1 g/day to about 12 grams per day, from about 5 grams per day to about 8 grams per ounce, from about 0.5 grams per ounce to about 4 grams per ounce, and from about 5 grams per ounce to about 2 grams per ounce. The peptide of the present invention can be used alone or in combination with a second drug. The 10th drug is a known antiviral agent such as an interferon-based therapeutic agent or another type of antiviral drug such as ribavirin. The second drug can be an anticancer agent, an antibacterial agent, or an antiviral agent. The antiviral agent can function at any step of the viral life cycle from the initial attachment to the invading cell. Thus, the added antiviral agent can interfere with attachment, fusion, invasion, travel, translation, viral multi-protein processing, viral genomic replication, viral particle assembly, detachment or budding. The antiviral agent can be an attachment inhibitor, an invasion inhibitor, a fusion inhibitor, a travel inhibitor, a replication inhibitor, a translation inhibitor, a protein treatment inhibitor, a release inhibitor, in other words, An inhibitor of viral function. The effective amount of the second drug will be determined by the second drug manufacturer's recommendation 20, as determined by the clinician, and will be determined by the PHYSICIAN'S DESK REFERENCE instructions and guidance on the dosage and dosage regimen and administration factors. The efficacy of the treatment may be as discussed above in the context of the viral infection sign or symptoms of the patient being monitored and the presence of viruses and viruses in the blood. The presence and/or presence of the virus, such as viral load, is determined. Methods used by the artisan include, but are not limited to, amplification of the chain reaction and amplification of the transcription medium. Pharmaceutical Compositions $+ In one embodiment, the invention provides a pharmaceutical composition comprising a peptide of the invention. To prepare such a pharmaceutical composition, the peptide of the present invention is synthesized or otherwise obtained, purified as needed or desired, and then dried and stabilized. The peptide can be adjusted to the appropriate concentration and then combined with other doses or therapeutically acceptable carriers. "Pharmaceutically acceptable" - the words mean that the carrier, diluent 10, excipients, and/or salts are compatible with the other ingredients in the formulation and are not deleterious to the recipient. Pharmaceutical formulations containing the therapeutic peptides of the present invention can be prepared by the well-known and readily available ingredients known in the art. For example, the peptide may be formulated using conventional excipients, diluents, or carriers, and may be formulated into tablets, elixirs, solutions, suspensions, powders, sprays, and the like. Examples of excipients, diluents, and carriers suitable for use in such formulations include buffers and fillers and bulking agents such as powders, celluloses, saccharides, mannitol, and derivatives of aspartic acid. Also included are binding agents such as methyl cellulose, methyl cellulose, propyl methyl cellulose and other cellulose derivatives, brown 20 alginate, gelatin, and polyvinylpyrrolidone. Wet lake agents such as glycerin may be included; disintegrating agents such as carbonated dance and carbonated carbonate. Delayed solubilizing agents such as rocky soil may also be included. A resorption accelerator such as a fourth ammonium compound may also be included. Surfactants such as lysine and stearic acid glycerin may also be included. Adsorbent carriers such as kaolin and bentonite may be added. Also 87 200837075 includes lubricants such as talc, calcium stearate and magnesium stearate, and solid polyethylene glycol. A preservative can also be added. The compositions of the present invention also include thickeners such as cellulose and/or cellulose derivatives. It also contains gums such as yellow gum, guar gum or carbo gum or gum arabic, or may additionally contain poly-5, bentonite and montmorillonite. For oral administration, the peptide may be in the form of a powder, a granule formulation, a solution, a suspension, an emulsion, or a resin which is used in a natural or synthetic polymer or from a chewing gum. The active peptide can also be in the form of a bolus, a sugar bait or a paste. The formulations may conveniently be presented in separate unit dosage forms as appropriate, and may be prepared by any of the methods well known to the pharmaceutical industry, including mixing the therapeutic agent with a liquid carrier, a solid base, a semisolid vehicle, a finely divided solid carrier, or a combination thereof. The step; then pouring or shaping the product into the desired drug delivery system if desired. The total active ingredient in these formulations is 0.1% by weight of the formulation. To 99.9% by weight. 15 Tablets or oval tablets containing the peptide of the present invention include buffers such as calcium carbonate, magnesium oxide and magnesium carbonate. Elliptical tablets and lozenges also include inactive ingredients such as cellulose, pregelatinized starch, ceria, hydroxypropyl decyl cellulose, magnesium stearate, microcrystalline cellulose, starch, talc, titanium dioxide, benzoic acid, Citric acid, corn house powder, mineral oil, polypropylene glycol, plantation, 20 stearic acid and so on. A hard or soft Mingsheng capsule containing at least one of the peptides of the present invention may contain inactive ingredients such as gelatin, microcrystalline cellulose, sodium sulfate, sodium sulphate, talc, titanium dioxide, etc., and liquid vehicles such as polyethylene. Glycols (PEG) and vegetable oils. In addition, enteric coated oval tablets containing one or more of the skins of the present invention are designed to counteract the disintegration of the stomach and dissolve in the more neutral to alkaline environment of the duodenum of 88 200837075. The therapeutic peptide of the present invention can also be administered orally by sustained release. In this case, the peptide of the present invention can be coated, microencapsulated (refer to WO 94/07529 and U.S. Patent No. 4,962, 〇91) or placed in a continuous delivery device. 5 A sustained release formulation can be designed to release the active peptide over a period of time in a particular part of the gut or respiratory tract. The coating, envelope, and protective substrate can be made, for example, from polymeric materials such as polylactide-glycolate, vesicles, microemulsions, microparticles, nanoparticles, or waxes. The coating, envelope, and protective substrate can be used to coat indwelling devices such as stents, catheters, peritoneal dialysis tubing, drainage devices, and the like. The therapeutic peptide of the present invention can also be formulated as an elixir or a solution conveniently, in a sedative manner, or as a solution suitable for parenteral administration, for example, by intramuscular, subcutaneous, intra-abdominal or intravenous routes. The pharmaceutical formulations of the therapeutic peptides of the present invention may also be in the form of an aqueous or non-aqueous solution or dispersion formulation or may additionally be in the form of an emulsion or suspension or ointment. This / 〇 胜 胜 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调 可调. As noted above, a preservative can be added to assist in maintaining the shelf life of the dosage form. The quinone component of the active peptide may be formed as a suspension, solution, or remedy for an oily or aqueous vehicle; it may contain a formulation such as a suspending agent, a stabilizer, and/or dispersion. Alternatively, the active ingredient and the other ingredients may be in the form of a powder, either by sterilizing the sterile solid or by lyophilizing the solution to obtain a powder form for use prior to use using a field vehicle such as sterile pyrogen free water. 200837075 5 10 Such formulations may contain pharmaceutically acceptable carriers, vehicles, and adjuvants which are well known in the art. The solution may be prepared using one or more organic solvents which are physiologically acceptable, and the organic solvent is selected from the following solvents other than water i, such as acetone, ethanol, isopropanol, glycol ethers such as Products sold under the trade name DGwanGl, polyglycols and polyethylene glycols, CrC4 money for short-chain acids, lactic acid or isopropyl lactate, fatty acids, triglycerides, etc. Products sold under the name "Mig (4)), isopropyl myristate, animal oil, mineral oil, and vegetable oils and polyoxyalkylenes. If desired, an adjuvant selected from the group consisting of antioxidants, surfactants, other preservatives, molding agents, keratolytic or cosmetic decomposers, perfumes, flavoring agents, and colorants may be added. An antioxidant such as a tert-butylhydroquinone butyl group, a butyl group, a butyl group, a hydrazine, and a bamboo organism can be added. In several embodiments, the peptide is formulated as a microbicide, either by topical administration or by administration to mucosal surfaces such as the vagina, rectum, eye, nose, and mouth. For topical administration, the therapeutic agent can be applied directly to the target area as is known in the art. The main dosage forms for topical administration include, for example, creams, emulsions, sizing agents, dispersing or microemulsions, thickening to a greater or lesser extent of 20 lotions, soaked pads, ointments or sticks. , spray formulations (such as sprays or foaming agents), soaps, detergents, lotions or soap cakes. Thus, in one embodiment, the peptide of the present invention can be formulated into a topical vaginal cream or microbicide. Other convenient dosage forms for use in this application include wound dressings, coated bandages or other polymeric coverings, ointments, creams 90 200837075, lotions, pastes, gels, sprays, and aerosols Agent. Thus, the therapeutic peptides of the present invention can be delivered through transdermal drug delivery patches or bandages. Alternatively, the peptide can be formulated as part of an adhesive polymer such as a polyacrylate or an acrylic/vinyl acetate copolymer. For long-term administration, it may be desirable to use microporous and/or breathable backing laminates, thereby reducing hydration or maceration of the skin. The backing layer can have any suitable thickness and will provide the desired protection and support functions. Suitable thicknesses typically range from about 10 microns to about 200 microns. Ointments and creams may, for example, be formulated with an appropriate thickening and/or gelling agent 10 in association with an aqueous base or an oily base. Lotions may be formulated with aqueous bases or oily bases which usually also contain one or more emulsifiers, stabilizers, dispersing agents, suspending agents, thickening agents, or coloring agents. The active peptide can also be delivered by ion electrophoresis, for example, as disclosed in U.S. Patent Nos. 4,140,122, 4,383,529, or 4,051,842. The weight percent of the therapeutic agent 15 of the present invention present in the topical formulation will be determined by a number of factors, typically from 0.01% to 95%, typically from 0.1% to 85% by weight, based on the total weight of the formulation. Drops such as ophthalmic drops or nasal drops may be formulated with one or more therapeutic peptides on an aqueous or non-aqueous base, and the drops also comprise one or more dispersing agents, solubilizing agents, or suspending agents. The liquid spray is conveniently delivered by a pressurized pack 2 cartridge. The drops can be delivered by a simple ophthalmic drop-capped vial or by a plastic bottle suitable for delivering the liquid contents drop-wise, delivered through a specially shaped closure. The therapeutic peptide can be further formulated for topical administration to the mouth or throat. For example, the active ingredient may be formulated into a diamond ingot further comprising a bridge-based base agent 91 200837075, which is often a sugar and acacia gum or a tragacanth; the soft capsule comprises a composition in an inert base such as gelatin and glycerin or sucrose. And acacia gum; and mouthwash comprising the composition of the invention in a suitable liquid carrier. The pharmaceutical formulations of the present invention comprise a pharmaceutically acceptable carrier, a diluent 5, a solubilizing agent, or an emulsifier and a salt of the type available in the art as an optional ingredient. Examples of such materials include physiological saline solutions such as physiologically buffered saline solutions and water. Specific non-limiting examples of carriers and/or diluents which may be used in the pharmaceutical formulations of the present invention include water and physiologically acceptable buffered saline solutions such as phosphate buffered saline solution pH 7.0-8.0. The peptide of the present invention is also administered to the respiratory tract. Thus the invention also provides aerosol pharmaceutical formulations and dosage forms for use in the methods of the invention. Generally, such dosage forms comprise at least one agent of the invention which is effective in treating or preventing the clinical symptoms of a viral infection. Any statistically significant attenuation of one or more of the symptoms of infection when treated in accordance with the methods of the present invention is considered to be a treatment for infections within the scope of the present invention. Alternatively, the composition may be in the form of a dry powder for administration by inhalation or insufflation, for example, a powder mixture of a therapeutic agent and a suitable powder base such as lactose or starch. The powder composition may be contained in a unit dosage form 20, for example, in a capsule or cartridge, or in a gelatin or blister pack, for example... from which the powder may be administered by means of an inhaler, an insufflator or a metered dose inhaler. (eg reference to pressurized metered dose inhalers (MDI) and dry powder inhalers as disclosed in Newman, SP, Aerosols and Lungs, Clarke, SW· and Davia, D. Ed., pp. 197-224, Butterworths , London, UK, 1984). 92 200837075 The therapeutic peptide of the present invention can also be administered in aqueous solution when administered in an aerosol or inhaled dosage form. Thus, other aerosol pharmaceutical formulations, for example, comprising a physiologically acceptable buffered aqueous saline solution containing from about 1 mg/ml to about 100 mg/ml are specific for the indication or disease to be treated 5 or A variety of peptides of the invention. Dry peptides which are finely divided solid peptides or nucleic acid particles which are not dissolved or suspended in a liquid can also be used in the practice of the present invention. The peptide of the present invention can be formulated as a dusting powder comprising finely divided particles having an average particle diameter of from about 1 micrometer to 5 micrometers and further from 2 micrometers to 3 micrometers. The method of making finely divided particles is prepared by pulverization and screening filtration using techniques well known in the art. The granules may be administered by inhalation of a predetermined amount of finely divided material, which may also be in powder form. It is to be understood that the unit content of the active ingredient contained in the aerosol of one of the dosage forms does not necessarily constitute an effective amount to treat a particular infection, indication or disease, as a plurality of dosage units can be administered to achieve the desired effective amount. In addition, an effective amount can be achieved using less than one of the dosage forms of the dosage form or a series of administrations. The therapeutic peptide of the present invention can be conveniently delivered by nebulizer or pressurized pack or other device for conveniently delivering a gas spray by administering it to the upper respiratory tract (nasal tract) or lower respiratory tract by inhalation. The pressurized pack may comprise a suitable propellant such as dichlorofluorocarbon, trifluorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or its appropriate gas. In the case of a pressurized aerosol, the metered dose can be delivered via a set valve to determine the dosage unit. Nebulizers include, but are not limited to, those described in U.S. Patent Nos. 4,624,251, 3,703,173, 3,561,444, and 4,635,627. The aerosol delivery system disclosed herein is available from a variety of commercial sources including Fisons Corporation (Bedford, MA) and 93 200837075 Schering Corp. (New Jersey) Cannix) and American Pharmoseal Co. (California, California). For intranasal administration, the therapeutic agent can be administered through nasal drops, liquid sprays such as through plastic bottle sprayers or metered dose inhalers. Typical sprayers are Mistometer (Wintrop) and Medihaler (Rike®). The therapeutic peptides of the invention may be combined with one or more Known therapeutic agents, such as pain relieving agents; antiviral agents such as anti-HBV agents, anti-HCV agents (HCV inhibitors, HCV protease inhibitors) or anti-therapeutic agents; antibacterial agents; 10 anticancer agents; anti-inflammatory agents ; an antihistamine; a bronchodilator, and a suitable combination thereof, whether used in the condition or in other conditions. Various compositions and articles of manufacture In one embodiment, the invention provides a composition comprising a peptide of the invention. A pharmaceutical composition used to control a manufactured article of a microbial infection. Such a device may be a useful device such as a vaginal ring, a condom, a bandage or the like. The device contains an effective amount of a pharmaceutical composition for controlling viral infection. The kit can be packaged in conjunction with an indication to control infection using a pharmaceutical composition. The composition of the composition comprises a therapeutically effective amount of at least one of the skins of the present invention to control viral infection. The kit may also be a container or filter unit for collection, processing or storage of a biological sample containing a peptide of the invention. The container includes, but is not limited to, a capillary tube, a vacuum container, a blood or other body fluid collection bag, a cannula, a catheter. The unit can be part of any device, such as a catheter for biological fluid collection. Furthermore, the peptide of the invention can also be attached to or covalently attached to the article 94 200837075, such as a container or filter unit. Thus, when the article is manufactured When the material is decanted or passed through the article, the material will not retain the peptide of substantial quality. However, the peptide is adsorbed or covalently attached to the manufactured article to kill the virus or prevent the spread of the virus' Controlling viral infections. Thus, for example, the peptide of the present invention can be incorporated into a filtration unit of a biological fluid collection conduit or container, or can be added to a collection trough to remove or deactivate a biological sample that may be present in the collected biological sample. Viral particles, thereby preventing the spread of the disease. The invention also provides a peptide comprising the invention and one or more clinically useful agents such as organisms The composition of the fixative. Biological stabilizers include, but are not limited to, 10 anticoagulants, preservatives, and protease inhibitors. Anticoagulants include, but are not limited to, oxalate, EDTA, and lemon. Acid salts and heparin. Preservatives include, but are not limited to, boric acid, sodium formate and sodium borate. Protease inhibitors include dipeptide-peptidase as inhibitors. Compositions comprising the peptides of the present invention and bio-stabilizers may be included in the collection. Containers for the collection, handling or storage of containers such as capillaries, vacuum containers, blood or other body fluid collection bags, cannulas, catheters or any other biological sample. The invention also provides for the inclusion of the peptides of the invention and in the laboratory The composition of the biological sample such as blood, semen or other body fluids of the recipient mammal is analyzed or desired. For example, the peptide of the present invention can be mixed with blood prior to laboratory treatment and / 20 or round of blood. In another embodiment, the peptide of the present invention may comprise a physiological medium for storing and transporting biological tissue, including transplanted tissue. Thus, for example, liver, heart, kidney, and other tissues can be immersed in a medium containing the peptide to inhibit viral infection to the recipient of the transplant. 95 200837075 The invention is further illustrated by the following non-limiting examples. EXAMPLES Example 1: Methods Cell Culture and Reagents: Huh-7 and Huh-7.5.1 cell lines are described by Zhong, J-5 et al., Proc Natl Acad Sci USA 102, 9294-9 (2005).

HeLa, Hep2, LLC-MK2, MRC-5, MA-104, Vero and MDBK cell lines were obtained from the American Type Culture Collection. All cells were maintained in DMEM supplemented with 10% fetal bovine serum (FCS), 10 mM Hepes buffer, 100 units/ml penicillin (penicillin), 100 mg / 10 ml streptomycin (streptomycin) And ΙΟΟμΜ non-essential amino acid (Invitrogen) at 5% CO 2 . Peptide synthesis: using L-amino acid or D-amino acid, using Amino® methoxycarbonyl (Fmoc) chemistry by A&A Lab, LLC (San Diego, CA), Highly purified peptide (> 95% purity) was synthesized by Mimotopes Pty Ltd (Clayton Victoria, Australia 15) or at the Scripps Research Institute. HCV manufacturing and infection: Infective genotype 2a HCV strain JFH1 was used in this study (see Kato, T. et al., J Med Virol 64, 334-9 (2001)). JFH_1 plastid construct for preparation of virus preparation, 20 cell culture conditions for JFH-1 infection, and analysis procedures for detecting the manufacture of virus are described in

Zhong, J. et al., Proc Natl Acad Sci USA 102, 9294-9 (2005). Indirect immunofluorescence: Intracellular staining of proteins or peptides is performed as described by Zhong, J. et al., Proc Natl Acad Sci USA 102, 9294-9 (2005). The recombinant human monoclonal (IgGl) anti-E2 anti-system is provided by D. Burton (Ref. 96 200837075)

Zhong, J. et al., Proc Natl Acad Sci USA 102, 9294_9 (2〇05)), anti-NS5A rabbit multi-strain resistance system provided by Μ·Houghton (Reference

Zhong, J. et al., Proc Natl Acad Sci USA 102, 9294-9 (2005)), and rabbit anti-dimethylaminonaphthalene-based multi-drug resistance system purchased from Molecular Probes (Oregon) State of Juki, and diluted 1:400 for use. Huh-7 cells were fixed in 4% paraformaldehyde, stained with the best dilution of these previously assayed antibodies, followed by 1:1 fold dilution of Alexa555-conjugated anti-human igG and anti-rabbit IgG Secondary antibodies (Molecular Probes Inc., Juk, Oregon) were co-cultured. The nucleus was stained with Hoechst dye or by differential interference contrast (DIC). The cellular localization of the protein or peptide was analyzed by the Radiance 2100 Rainbow Laser Scanning Confocal Microscope (LSCM) (Bio-Rad, Hercules, CA). MTT cytotoxicity assay: The cytotoxic effect of the peptide is based on the manufacturer's instructions (MTT assay suite, catalog number 30-1010K, ATCC, Manassas, VA) on V[TT] Toxicity assay analysis. Briefly, a series of 2-fold diluted peptides were added to 5,000-10,000 cells in 96-well plates in 5% dextrose in water containing 5% DMSO or DMSO alone. After 72 hours at 37 ° C, a 1/1 volume of MTT solution (5 mg/ml in PBS) was added to each well to return the well plate to the incubator. After 2 hours, the medium was removed, 150 μl of DMSO was added to dissolve the f0Tmazan precipitate, and the well plate was shaken at 150 rpm for 10 minutes, and then the absorbance ratio was read at 570 nm. The peptide concentration, which reduced the 〇d reading by 50%, was designated as 5〇% lethal concentration (Lc5〇). In vivo cytotoxicity assay: To determine the cytotoxicity of peptides in mice 97 200837075 Potential, pure peptide (>95%) is dissolved in DMSO at a concentration of 50-100 mg/ml and then diluted in a non-class 5% glucose/water of toxin, 2 〇〇 microliter peptide solution or DMSO (5%) control solution intravenous (tail vein) injection into multiple groups of 7-9 week old C57BL/6J mice, the mice were called before injection. Heavy, and weighed on the 5th, 2nd, 4th, and 6th and 8th after the injection. The selected mice were injected with 〇5 mg of D-isomer three times per week, and every other week after the injection was monitored for one week. In vivo immunogenicity assay: To determine whether the peptide is immunogenic, the serum obtained by intravenous injection of 5 mg of D_isomer in mice at weekly intervals is tested for peptides using a peptide-specific ELISA. antibody. Briefly, i 10 μg D_Peptide 1 or PBS was applied to ImmimoPlates (Nalge Nunc Intl, Rochester, NY) at 4 °C to isolate Washed with pbs (wash buffer) containing 0.05% Tween 2 times; then blocked overnight at 4 °C with 5% FBSi5% non-fat dry milk; washed 4 times, then added a series of 2 The diluted mouse serum was diluted at 15 1:10 and incubated for 1 hour at room temperature. The wells were then washed 10 times and conjugated with twice horseradish peroxidase (HRP) goat anti-mouse 1 § <3 (1:25 〇〇〇 dilution) (Pierce, Illinois, Locke) Fu) Co-cultivation at room temperature, washing 10 times, developing with tetramethylbenzidine (Pierce) according to the manufacturer's instructions, measuring optical density at 650 nm, comparing 20 wells coated with peptide And the PBS-coated control wells, the peptide-coated wells were considered positive by comparison with the PBS-coated pores by more than 2 times the absorbance. The assay control group consisted of 1 μg of dimethylaminonaphthalenesulfonated D-peptide 1 and multiple anti-dimethylaminonaphthalene thiol antibodies (1:4 dilution) with rabbits. Co., Ltd., Jukin, Oregon) co-cultured with HRP-conjugated goat anti-rabbit IgG (1:25, 〇〇〇200837075 dilution) (Pierce, Rockford, Ill., IL); b) coated with 1 microgram of influenza hemagglutinin (HA) peptide (Sigma, St. Louis, MO) and mouse monoclonal anti-HA antibody (1:600 dilution) (West) Comar, St. Louis, Missouri, co-cultured the wells co-cultured with HRP-conjugated goat anti-rabbit IgG (1:25,000 dilution) (Pierce, Rockford, Ill.). Velocity Settling Ultracentrifugation: The settling velocity of native and peptide-treated infectious HCV particles is as previously described by the ultracentrifugation of the rate section to the continuous cane gradient test. Briefly, 100 microliters of the sample was plated in 5 ml of a pre-formed ° to 50% continuous sugar gradient and centrifuged at 200,000 X g for 1 hour at 4,000 X g in a sw60.Ti rotor at 4 C. After centrifugation, n fractions (400 microliters each) were collected from the top as analyzed for viral infectivity and hcv RNA content as described above. Intracellular HCV infectivity assay: Huh-7 cells were infected with jFH_i at an infection fold of 15 (m〇i) = 0.01. After 10 ,, the infected cells were washed 3 times with PBS, decomposed by trypsin, and resuspended in complete medium at a concentration of lxlO5 cells/ml. The cells were lysed in dry ice and 37 〇c water bath by 4 freeze-thaw cycles, and the cell debris was removed by centrifugation at I4,00 (^pm. The supernatant was assayed for the infectivity of the supernatant as described in the previous titration assay. It virus money: To determine whether the peptide of sputum can inhibit other virus infections, do not add the concentration of each peptide or DMS 至 to a powerful virus preparation (1-105 ffu or TdDW ml), and culture at 37t for at least 1 hour. And then added to the susceptible cells (unless stated). After 2 to 4 days of infection, by comparative evaluation (4) cell-induced (CPE), or as follows, immunostaining 99 200837075 color or using anti-corresponding viral proteins The antibody was evaluated by immunoassay. Since HBV is not infectious in vitro, the effect of peptide on the antigenicity of HBs was examined by quantitative ELISA analysis, such as Guidotti et al., J Virol 69, 6158-69 (1995). And; by quantitative PCR analysis to examine the effect of peptide on 5 HBV DNA content, as described by Thimme et al, j vir〇i 77, 68-76 (2003). Example 2: HCV peptide inhibits hepatitis C virus Infection example illustrates the antiviral peptide of the present invention Identification: Preparation of a peptide library of the 441 overlapping peptide of the hepatitis C virus polyp 10 white (SEQ ID NO: 1) covering the hepatitis C genotype la (H77), using a cell culture model of hepatitis C virus infection The inhibitory effect of the test on hepatitis C virus infection as described in the relevant application (u s. Ser. N〇. Π/541, 488). The peptide is about 18 amino acids long and has 11 overlapping amino acids. The peptide library was manufactured by the National Institutes of Health (NIH) AIDS Research and Reference Reagent Program (catalog number 762〇, Batch No. #1). To identify peptides showing antiviral activity against HCV infection, HCV assay analysis To select the peptide library. The peptide was reconstituted in 100% DMSO at a final concentration of ι〇mg/ml and stored in -2 (rc. peptide standby solution in complete DMEM containing hcV of 5 〇 lesion forming unit (ffu) The growth medium was diluted 1.200 to about 2 μM. The virus-peptide mixture was transferred to 5 1 cells, and the cell density was 8 cells per well in a 96-well plate. At 37. 〇 desorption 4 After the hour, 'remove the lion. The cells are washed 2 times, and the top is covered with microliters of fresh growth medium at 3rc. After 3 days of culture, the cells were fixed with paraformaldehyde, immunostained with anti-HCV non-structural protein NS5A antibody, and the number of HCV lesions was calculated under a fluorescence microscope. The results were obtained by inoculating virus and 〇·5% DMSO. However, the percentage (%) of the control lesions detected in the cells without the peptide is indicated. The results of these assays are shown in Figure 1 and the table below. Table 3: Inhibition of HCV infection

Number peptide sequence % pseudo-inhibition multiple > 10-fold 5-10 times 2-5 times SEQ ID NO: 6930 QIVGGVYLLPRRGPRLGV 45·2 2.2 氺 4 6937 QPGYPWPLYGNEGCGWAG 50.0 2.0 * 5 6938 LYGNEGCGWAGWLLSPRG 2.4 42.0 氺氺氺6 6939 GWAGWLLSPRGSRPSWGP 45.2 2.2 * 7 6951 IFLLALLSCLTVPASAYQ 2.4 42.0 *氺氺8 6957 DAILHTPGCVPCVREGNA 21.4 4.7 * 9 6962 LPTTQLRRHIDLLVGSAr 38.1 2.6 氺10 6963 RfflDLLVGSATLCSALYV 31.0 3.2 氺11 6964 GSATLCSALYVGDLCGSV 1.0 100.0 12 6965 ALYVGDLCGSVFLVGQLF 1.0 100.0 13 6975 MDMIAGAHWGVLAGIAY 2.4 42.0 氺氺氺14 6986 HINSTALNCNESLNTGWL 40.5 2.5 Shui 15 6987 NCNESLNTGWLAGLFYQH 35.7 2.8 Shui 16 6991 LASCRRLTDFAQGWGPIS 35.7 2.8 * 17 6992 TDFAQGWGPISYANGSGL 31.0 3.2 Shui 18 6993 GPISYANGSGLDERPYCW 23.8 4.2 Shui 19 6994 GSGLDERPYCWHYPPRPC 33.3 3.0 Shui 20 7005 WMNSTGFIXVCGAPPCVI 16.7 6.0 Shui Shui 21 7007 PCVIGGVGNNTLLCPTDC 33.3 3.0 * 22 7016 MYVGGVEHRLEAACNWTR 16·7 6.0 氺* 23 7026 YLYGVGSSIASWAIKWEY 2.4 42.0 氺氺氺24 70 27 SIASWADCWEYWLLFLL 40.5 2.5 氺25 7028 KWEYWLLFLLLADARVC 47.6 2.1 氺26 7031 WMMLLISQAEAALENLVI 4.8 21.0 氺氺氺27 7038 GAVYAFYGMWPLLLLLLA 19.0 5.3 氺28 101 200837075 Number 胜 序列 % % & & & & & & & & & & & & & & & & & 5 times SEQ JD NO: 7039 GMWPLLLLLLALPQRAYA 31.0 3.2 * 29 7052 TLVFDITKLLLAIFGPLW 1.0 100.0 *** 30 7725 VSTATQTFLATCIN 40.5 2.5 氺31 7078 ATQTFLATCINGVCWTVY 2.4 42.0 氺氺本32 7142 DSSVLCECYDAGCAWYEL 40.5 2.5 氺33 7146 AYMNTPGLPVCQDHLEFW 40.5 2.5 氺34 7148 LEFWEGVFTGLTHIDAHF 33.3 3.0氺 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 100.0 氺氺* 43 7209 DWICEVLSDFKTWLKAKL 2.4 42.0 氺氺氺44 7226 YVSGMTTDNLKCPCQIPS 38.1 2. 6 氺45 7740 SSGADTEDWCCSMS 42.9 2.3 氺46 7741 DTEDWCCSMSYSW 2.4 42.0 氺氺氺47 7270 SSGADTEDWCCSMSYSW 4.5 22.0 氺氺氺48 7742 DWCCSMSYSWTGAL 23.8 4.2 氺49 7304 TVTESDIRTEEAIYQCCD 35.7 2.8 氺50 7313 GNTLTCYIKARAACRAAG 45.2 2.2 * 51 7315 RAAGLQDCTMLVCGDDLV 50.0 2.0 氺52 7316 CTMLVCGDDLWICESAG 1.0 100.0 氺氺氺53 7317 DDLWICESAGVQEDAAS 26.2 3.8 氺54 7323 LELITSCSSNVSVAHDGA 42.9 2.3 氺55 7329 HTPVNSWLGNEMFAPTL 47.6 2.1 氺56 7331 APTLWARMILMTHFFSVL 45.2 2.2 * 57 7334 DQLEQALNCEIYGACYSI 28.6 3.5 * 58 7342 GVPPLRAWRHRARSVRAR 50.0 2.0 * 59 7343 WRHRARSVRARLLSRGGR 47.6 2.1 氺60 7350 GWFTAGYSGGDIYHSVSH 42.9 2.3 氺61

102 200837075 Number peptide sequence total % pseudo-suppression >10-孓10 2-5 m Analog multiples 14 4 41 441 peptide, 382 has no effect on HCV infection, or block infection less than 20% ( Not shown in Table 3). 41 peptides slightly inhibited HCV infection by about 2 to 5 times. The four peptides inhibit HCV infection by about 5 to 10 fold. 14 peptides inhibited ‘ 5 HCV infection by more than 1〇. In particular HCV infection was significantly inhibited (90-100%) by the peptides of SEQ ID NO: 6, 8, 12, 13, 14, 24, 27, 30, 32, 43, 44, 47, 48 and 53. No evidence of toxicity was detected when Huh-7.5.1 cells were co-cultured with these peptides. These results identify a peptide inhibitor that modifies or inhibits one or more steps in the viral life cycle. Further, 10 according to the present invention, these peptides can be used for an antiviral composition and a method for inhibiting HCV infection. Peptides that inhibit infection by more than 90% were selected for further analysis. The antiviral activity of these inhibitory peptides was verified by comparing the height of each peptide (20 μM) (>95%) to the purified preparation (infection) by comparison with the solvent control group (0.5% DMSO). M〇i 〇. 1) The ability to inhibit the proliferation of HCV 15 RNA in Huh-7.5.1 cells 72 hours later. In order to quantify the inhibitory effect of the peptide on HCV infection, RT-QPCR was used to quantify the intracellular HCV RNA content of the peptide-treated cells and untreated cells. The peptide backup solution and the DMSO solvent were diluted 1:100, and the supernatant of the alternate virus was mixed to obtain a final peptide concentration of 18 μM and a final DMSO concentration of 0.5%. The virus-peptide mixture and the 20 virus-DMSO mixture are then used to infect at a multiple of infection (moi) 0.1

Huh-7_5.1 cells. After 3 days of culture at 37 ° C, the cells were washed, dissolved, and the total cellular RNA was separated. 103 200837075 After adding 5 μg of yeast tRNA as a carrier per 500 μl sample, total cellular RNA was isolated by the thiocyanate method using standard protocols. For example, Cheng et al., Proc Natl Acad Sci USA 103, 8499-504 (2006) performed reverse recording and quantitative real-time PCR (RT-QPCR). 5 The HCV and glyceraldehyde_3_phosphate dehydrogenase (GAPDH) transcript levels were determined as compared to a standard curve consisting of serial dilutions of one of the plastids containing the HCV JFH-1 cDNA or the human GAPDH gene. After normalizing the GAPDH mRNA content of the cells, the relative HCV RNA content of the infected cells was determined. The detection limit of the RT-QPCR protocol is approximately one set of HCV 10 RNA per 1000 cells, using RT-QPCR, using primer 5, _TCTGCGGAACCGGTGAGTA-3' (message, SEQ ID NO: 89) and 5,-TCAGGCAGTACCACAAGGC-3, (Anti-message, SEQ ID NO: 90) The HCV RNA transcript content was determined and normalized to the cell 15 GAPDH content. The inhibitory activity was measured by comparing the concentration of intracellular HCV RNA after the peptide-treated inoculum and the solvent-treated inoculum were normalized. The results are summarized in the table below. 104 200837075 Table 4: Inhibition of peptide peptide HCV RNA inhibition fold peptide peptide amino acid sequence SEQ m 24h 25μΜ 72 h 25_ 1 NS5A 1975 membrane anchor SWLRDIWDWICEVLSDFK 43 1,860 245,6 58 2 NS5B 2731 catalytic functional site CTMLVCGDDLWICESAG 53 86 40 3 NS5A/5B 2413 ''BILN2061' SSGADTEDWCCSMSYSW 48 75 49 4 Ε2/Ρ7 736 WMMLLISQAEAALENLVI 27 27 40 5 Ε1 267 putative fusion peptide GSATLCSALYVGDLCGSV 12 26 27 6 NS2 883 TLVFDnXLLLAlFGPLW 30 24 16 7 Ε1 274 putative fusion peptide ALYVGDLCGSVFLVGQLF 13 20 11 8 Core/El 176 signal peptide cleavage IFLLALLSCLTVPASAYQ 8 18 7 9 E1344 MDMIAGAHWGVLAGIAY 14 10 22 10 Core 85 LYGNEGCGWAGWLLSPRG 6 9 11 11 E2 701 YLYGVGSSIASWAIKWEY 24 9 7 12 NS3 1065 ATQTFLArdNGVCWTVY 32 8 5 13 NS5A1982 Membrane anchor DWICEVLSDFKTWLKAKL 44 7 2

Based on the infectious class, the most active peptide is relabeled with a numerical value to reflect the position of the peptide in the infectious class, as shown in Table 5. Thus derived from the NS5AN-terminal peptide 1 (the most potent peptide inhibitor of SEQ ID NO: 43@HCV, inhibits the proliferation of the virus by more than 4 times. The peptide 1 contains the amphipathic helix of the HCV NS5A protein. The terminal membrane anchors the residues of the functional part 3-21. However, there are peptides of SEQ ID NO: 6, 8, 12, 13, 14, 24, 27, 30, 1〇32, 44, 47, 48 and 53. The cell culture research model 105 of the Hepatitis C virus infection described in the related application (US·Ser. Νο. 11/541, 488) can also strongly inhibit HCV by measuring the type 20087777575. Other peptides have a good inhibitory effect on HCV infection. The HCV-derived synthetic peptides, which are potent inhibitors of peptides, are derived from both the structural and non-structural regions of the HCV polyprotein. Example 3: Analysis of the N-terminal and C-terminal truncated peptides 1 In order to define the peptides# 1 (SEQ ID NO: 43) antiviral activity, using a lesion reduction assay and measuring the decrease in intracellular HCV RNA as described to analyze a series of peptide N-terminal truncations and C-terminal truncated antiviral Highly purified peptide (>95% purity) was used for these studies. All peptides were 10 systems using fluorenylmethoxycarbonyl (Fm) Oc) Chemistry is synthesized by pre-loaded wang resin A&A Laboratories (San Diego, Calif.) at the Symphony multi-peptide synthesizer (protein Technologies Inc, The peptide was synthesized in Tucson, Arizona. The crude peptide was then purified and analyzed by a reversed Gilson HPLC system (Gilson 15 Division, Midotown, Wisconsin). C18 column (Grace Vydac, California), with a bead size of 20 mm and a length of 250 mm. The solvent system is water and B | solvent system is 5% to 70% The linear gradient was subjected to 30 minutes. Mass spectrometry was performed by a PE Sciex API-100 mass spectrometer. This confirmed the molecular weight of the synthetic peptide. The peptide concentration was the use of a chromophore residue (tryptophan or Determination of the extinction coefficient of tyrosine, where tryptophan = 5560 AU / millimoles / ml and tyrosine = 1200 AU / millimoles / ml. Calculated using the formula: mg peptide / ml = (A280 X DF X MW) / e, where A280 is a solution of 280 nm in a 1 cm light test tube The actual absorbance ratio, DF is the dilution factor, MW is the peptide molecule 106 200837075, and e is the molar extinction coefficient of each chromophore group at 280 nm. The results are summarized in the table below, and the results show that there is a C-terminal truncation 1 The peptide to 4 amino acid residues still retains antiviral activity. The removal of as few as 2 amino acid groups from the N-terminus destroys antiviral activity. 5 Table 5: Anti-HCV activity of the truncated variant of peptide 1

SEQ m peptide

SWLRDIWDWICEVLSDFK

SWLRDIWDWICEVLSD

SWLRDIWDWICEVL

SWLRDIWDWICE

SWLRDIWDWI

SWLRDIWD

LRDIWDWICEVLSDFK

DIWDWICEVLSDFK

WDW1CEVLSDFK

WICEVLSDFK

CEVLSDFK pseudo-simulation (Mock) lesion reduction (72h) inhibition ratio of HCV RNA (24h) inhibition ratio of HCV RNA (72h) 47697.9 301779.7 9852.6 234207.1 20274.7 237172.4 0.8 0.5 0.8 0.6 1.3 1.0 0.7 0.6 1.0 0.7 2.2 1.4 1.1 1.0 1.1 0.7 43 19.5μΜ 0 94 18.4μΜ 0 92 23.6μΜ 0 104 21.4μΜ 107 105 25·6μΜ 65 106 24.7μΜ 58 107 18.6μΜ 125 108 27.1 μΜ 125 109 24·7μΜ 38 110 27·5μΜ 45 111 ΝΑ 53 51 Example 4: The peptide prevents the initiation of HCV infection and suppresses the established infection based on the virucidal activity of peptide 1, and the following experiment was conducted to determine whether peptide 1 completely and permanently inhibits the establishment of HCV infection. Huh-7.5.1 cells were subjected to HCV (moi 0.1) and peptide 1 (ΐ8 μΜ) or 0.5% DMSO as controls. After 4 hours of adsorption at 37 ° C, the virus-peptide inoculum and the virus-DMSO inoculum were removed, the cells were washed twice, and 12 μL of fresh growth medium was placed over the cells, and incubated at 37 °C. At the indicated time point, total cells 107 200837075 RNA were isolated and assayed for HCV RNA concentration. As shown in Figure 2A, in contrast to the observed spread of the virus in the control culture, the intracellular viral RNA gradually decreased in the cells infected with the virus that was incubated with the peptide 1 before inoculation (less than a set of duplicate gossip). 〇〇 cells), maintenance can not be detected for at least 11 曰 time. These five results suggest that peptide 1 prevents the onset and spread of HCV infection by virucidal activity. In order to determine whether peptide 1 can end the ongoing HCV infection, Shengyue Tai 1 is 3 days after virus inoculation, and when 10% of the cells are positive for HCV E2, it is added to the infected cells and maintained in the medium, each time Cell division during filling. 10 More specifically, Huh-7 cells were infected with HCV (moi 0.1). On the third day after infection, when about 10% of the cells were infected (HCV E2 positive by immunostaining), peptide 1 (18 μΜ) or 0.5% DMSO was added, and the cells were inoculated at 37 ° C to reach the fusion every 3 - Split on the 4th 1:6. Each time the peptide and DMSO were added, the cells were divided, and total cellular RNA was isolated at this time point, and the content of HCV RNA transcription was determined. As shown in Figure 2B, although intracellular HCV RNA increased in DMSO-treated cells, L-form and D-form peptide 1 (SEQ ID NO: 43) suddenly stopped viral proliferation, and viral RNA slowed down to 45th. At least one set of copies per 1000 cells. Importantly, HCV RNA remains negative for at least 5 days after withdrawal of the peptide (not shown). These results suggest that the peptide 20 1 not only prevents HCV infection by destroying the virus, but also blocks the cell-to-cell spread, while also terminating the ongoing HCV infection in the dividing cell culture.

In order to determine whether peptide 1 can suppress infection with undivided cells with established infection, add to the 15th day after infection, when more than 9〇% of cells HCV 108 200837075 E2-positive (not shown), D form wins Peptide 1 and DMSO were added to highly differentiated Huh-7 cells that stopped growth (see Sainz et al, J. Virol. 80 10253-7 (2006)). Specifically, Huh-7 cells were treated with 1% DMS0 for 10 曰 to induce differentiation and growth arrest, at which time HCV was infected with moi=〇.〇i. 5 On the 15th day after infection, when more than 90% of the cells were HCVE2-positive, the L form of the peptide 1 and the D form (18 μΜ) were added to the complete growth medium. Each medium was replaced with a medium containing a fresh peptide. At the indicated time point, total cellular RNA was isolated and the HCV RNA content was measured. For comparison, infected cells were treated with 100 units/ml recombinant human IFNa (PBL Biomedical Laboratories, New Jersey Pitz 10 Kawe) and replaced daily as described above. As shown in Figure 2C, by day 5, the peptide 1 D form suppressed intracellular HCV RNA to the same level as IFNa, and even more inhibited than IFNa on the 15th and 20th day of treatment (> 95%). The results indicate that peptide 1 strongly inhibits persistent HCV infection in cells with impaired growth. 15 These results also suggest that in addition to extracellular virucidal activity, peptide 1

It is also possible to activate HCV intracellularly. To test this hypothesis, the peptide i was evaluated to determine if it could enter the cell and cause instability of the intracellular viral particles. In order to determine whether the peptide can substantially enter the cell, the fluorescent form of the peptide containing the dimethylaminonaphthalenesulfonyl group at its N-terminus is diluted to a final concentration of 18 μΜ in complete medium, and at 37 ° C with Huh. -7 cells were cultured for 4 hours and then washed 5 times with pbS. The treated cells were then fixed with 4% paraformaldehyde and immunostained using a polyclonal antibody against diammonium naphthalene xanthine (Molecular Probes Inc., Juk, Oregon) for analysis by confocal fluorescence microscopy. . As shown in Fig. 2D, the fluorescent form of the peptide 1 containing the dimethylaminonaphthalenesulfonyl group at its N-terminus is effective. 109 200837075 The Huh-7 cells are introduced into the granule structure of the cytoplasm. Importantly, when the infected cells were cultured for 6 hours with the D-isomer of peptide 1, the number of intracellular infectious HCV particles was reduced by a factor of 3 (Fig. 2E), while intracellular HCV RNA (not shown) did not have any Variety. Huh-7 cells infected with JFH-1 at moi 0.01 for 5 days were washed 4 times and treated with peptide 1 D-isomer (18 μM) or DMSO (0.5%). After 6 hours of culture, intracellular HCV infectivity, extracellular HCV infectivity, and cellular HCV RNA content were measured. The results suggest that in addition to the extracellular viricidal activity, peptide 1 can enter the cell and deactivate the intracellular virus, although it is ineffective and does not block the replication of HCV. 10 To determine the intermediate effective concentration (EC5〇) of peptide #1, the peptide backup solution (3·6 rnM in DMSO) was diluted 2-fold in DMSO-series. A whole portion of the peptide obtained by each dilution was diluted 1:100 in complete growth medium and mixed with an equal volume of viral supernatant. The virus-peptide mixture was then used to infect Huh-7.5.1 cells (ΜΟΙ=0·1). After adsorption at 37 ° C for 4 hours, the virus-peptide was inoculated 15 . The cells were washed twice, 120 μl of fresh growth medium was placed above, and cultured at 37 ° C for 3 days. The cells are lysed and subjected to RNA analysis. The HCV RNA transcript content was measured by RT-QPCR in time and normalized to the cellular GAPDH content. The inhibition of HCV infection was determined by comparing the intracellular HCV RAN transcript between the peptide treatment group and the solvent control group. The results (Fig. 2F-G) show EC5 of peptide #1 under these 20 pieces. It is about 300 nM. Example 5: Antiviral activity assay of peptide #1 (SEQ ID NO: 43) To determine the antiviral activity of peptide #1, it was tested to prevent viral RNA cells from being infective cells. The ability of cells to bind/attach/intake. Huh-7.5.1 cells were seeded in 96-well plates at 8 〇〇〇 cells/well. 110 200837075 16 j, after the presence of the skin with or without Handu ΜμΜ, the cells were co-cultured with gey at MOI=〇.l. After 3 η: adsorption for 4 hours, the virus _ peptide inoculum was removed. The cells were washed twice, dissolved, and subjected to RNA analysis. The HCV RNA transcript was determined by a quantitative quantitative polymerase chain reaction (RT-QPCR) assay and quantified to the cellular GAPDH content. The inhibitory activity was quantified by comparing the amount of HCV RNA bound to the cells in the cells exposed to the virus-peptide inoculum to the virus-DMSO control group. The results (Figure 3) indicate that Shengxin (and peptide 2, the overlapping peptide η significantly blocked the binding/attachment/uptake of the virus, and no other peptides were active at this concentration. 10 To further define Active machine rotation, relative to the addition of inoculum

Between, peptide #1 was added to the cells at different times. In order to determine the mechanism of action, in other words, in order to determine whether the inhibitory peptide at the concentration of the virus or the cell or both functions, the L-isomer of peptide 1 is added to the different time relative to the inoculation according to the following protocol. Virus or added to the cells. Huh-7.5.1 fine cells were seeded at 8000 cells/well in 96-well plates and infected with 800 ffu/well HCV the next day. The peptide was added at a final concentration of 18 μM under the following conditions: (1) Pre-inoculation: Before the virus infection, the peptide was added to the cells at 37 ° C for 4 hours, followed by washing with the growth medium 4 times; (2) simultaneous inoculation: win The peptide was added to the cells together with the virus for 4 hours. At this time, the cells were washed as above and supplemented with the complete culture of the thiol group without the peptide; (3) post-inoculation: the cells were infected for 4 hours, at which time the virus was removed and the peptide was added. , stayed on the cells and went through the entire experiment without washing off. At 24 hours and 72 hours after infection, the cells were lysed, and the HCV RNA concentration was determined by rt-QPCR in time and normalized to the cellular GAPDH content. As shown in Fig. 4B, peptide 1 has a strong inhibitory effect of 111 200837075 if it is added to the cells together with the virus; if it is added to the cells 4 hours after infection, it is far less effective, if it is added to the cells for 4 hours and Removal of the virus before it is added has no inhibitory effect at all. The results suggest that peptide 1 blocks very early steps in the HCV life cycle or has a virucidal effect on HCV. 5 In order to distinguish these alternative changes, HCV viruciens assay was performed in which the viral supernatant was pretreated with L-isomer or D-isomer or DMSO for 1 hour, and then the HCV RNA content remaining in the supernatant was determined. And the appeal. In order to determine whether the peptide has virucidal activity, the peptide is diluted to a concentration of 18 μM in a complete growth medium containing 5χ1〇5 ffu/, and cultured at 37 ° C for 10 hours, at which time the virus-peptide mixture is analyzed. Total HCV RNA and infectivity were compared to the comparatively prepared virus _DMS 〇 control group. The HCV RNA content was quantified to GAPDH RNA concentration by timely rt_QPCR assay, and the GAPDH RNA was added to RNA samples to control RNA extraction efficiency. The HCV infectivity was determined by diluting each 25 μl sample in the growth medium by a factor of 200 (ie, below the inhibitory concentration of the diluted peptide), by co-cultivating the diluted sample with Huh-7.5.1 cells, 3 The number of HCVE2-positive lesions was calculated in the future to determine residual infectivity. As shown in Fig. 4C, compared with the DMS 〇 control group, the peptide was cultured within the sputum hour, and the total HCV RNA content and infectivity were reduced by 1 及 and 20 10,000, respectively. The results suggest that peptide 1 is virucidal to HCV. To verify that this interpretation was performed by rate segment ultracentrifugation on a continuous sucrose gradient, the sedimentation rate of untreated, peptide-treated and Dms(R) treated virus particles was measured. Briefly, the L- and D-isomers (18 μM) of peptide 1 were added to separate virus fractions containing 1χ1〇5 ffu/ml HCV and cultured for 3 hours at 3rt 112 200837075. The third whole virus was co-cultured with 0.5% DMSO as a control. After the cultivation, 100 μl of the sample was analyzed by speed sedimentation ultracentrifugation as described in the method. Twelve selections (400 microliters each) were collected from the top to analyze viral infection and HCV RNA content. As shown in Figure 4D, the DMSO control sample showed a unique viral RNA peak that was substantially eliminated by the L- and D-isomers. As expected, the appeal of all selected points was also completely destroyed. In a nutshell, these results strongly suggest that the structure of the peptide that destroys HCV is intact and is also virulent for HCV. 10 Since the peptide 1 is derived from the membrane anchoring function site of NS5A, it is predicted to be an amphipathic α-helix in the same plane, which can cause metastasis and destruction of viral particles via the membrane penetrating the HCV virus particles. To test this hypothesis, a liposome dye release assay was performed. The liposome dye release assay involves the co-culture of cholesterol-phospholipid vesicles of encapsulated fluorochrome with peptide or DMSO and measuring the release of fluorescein. The preparation of microlipids (large monolayer vesicles, LUV) was as follows. a mixture containing 28 mg of total lipid (12 mM), consisting of 10 parts of 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC), 11 parts of 1,2-dipal醯-sn_glycerol-3_phosphocholine (DOPC), 1 part 1-palmitoyl-2-oleyl-sn-glycerol 20 _3_[phosphoric acid] (P〇PS), and 6 parts Cholesterol (Avanti Polar Lipids, Inc., Alabaster, Alabama), this mixture is dissolved in 1 ml of chloroform, 1 ml of ether and 2 ml of Rhodamine B (100 mM) At 10 mM Hai Pei Si, pH 7.2; Sufu B (Sulfo B), Molecular Probes Inc., Jukin, Oregon). The mixture was processed in a Branson 113 200837075 (Brans〇n) 2210 water bath ultrasonic oscillator (Danbury, Connecticut) at 4 C for 1 minute with an ultrasonic disk. After removal of the organic solvent in the Buchi Rotvapor R-114 (Labortechnik AG, Flavi, Switzerland), the lipid was resuspended in an additional 2 ml of Sufu 5 B 'Using 逑Your extruder (Mini-Extruder) (Afangdi Polar Lips, Alabaster, Alabama), repeated extrusion through a stack of 8 μm, 〇·4 μm and 〇·2 μm polycarbonate A membrane filter was used to screen the resulting lipid vesicles. Separation of the succulent scum of the sulphate was separated from the unentrained sulphate on the G-25 column. Aminco-Bowman Series 2 Cold Light 10 Light 5 Jin Yi (Thermo Electron Corporation, Wattham, MA) was used for dye release assay analysis. Twelve microliters of the vesicles were diluted in 1 ml of Haipeisi buffer to a final concentration of 120 μM in a stirred light test tube at room temperature. The sample was lasered at 535 nm wavelength and monitored at 585 nm. After 6 seconds of equilibration, start at ι〇μΜ 15 concentration, add peptide (10 μl) to the light tube, and monitor the kinetics of membrane destruction by increasing the fluorescence of Sufford. The percentage of Sufu sputum released by the addition is calculated by the following equation: % released Sufu Β = 100 * (F-FoWF j where F is the fluorescence intensity observed in the presence of the peptide, F() is the victory Peptide Add 20 plus the base fluorescence intensity, which is the fluorescence intensity corresponding to 100% Sufford beta release obtained by adding 25 μl of 10% Tdton X_100. As shown in Figure 4E, 'When DMSO In the absence of activity, the L- and D-isomers (1〇μΜ) of peptide 1 are immediately infiltrated with 70% liposome. This effect has a dose dependency until it is as low as 160 ηΜ, at this point, 20% micro Lipid penetration (not shown in Figure 114, 200837075). These results suggest that the virucidal effect of peptide 1 reflects its ability to penetrate cholesterol-rich phospholipid membranes. Example 6 · In vitro and in vivo In the non-cytotoxic concentration, peptide L and D-forms inhibit HCV infection. 5 The peptide consisting of L-amino acid is sensitive to the decomposition of the protein, shortening its lifespan and thus reducing its biological activity. It is possible to determine whether a specific peptide-to-viral protein interaction can mediate antiviral activity, and peptide 1 uses a full D-amine. The base acid is synthesized and purified to greater than 95% homogeneity, and its antiviral activity and serum stability are compared with the L-form version of the peptide #1 by a formulation having a similar purity of 10. L-type peptide and D- The peptide was diluted 1:100 in complete growth medium (10% FBS) and mixed with an equal volume of virus supernatant. 0 In addition, in order to compare the serum stability of L-form and D-type peptide, the diluted peptide was mixed. The virus supernatant was incubated at 37 ° C for 1 hour, 2 hours and 4 hours 15 hours. The virus-peptide mixture was then used to infect Huh-7.5.1 cells (ΜΟΙ = 0.1). Adsorption at 37 ° C 4 After the hour, the virus-peptide inoculum was removed. The cells were washed twice, and 120 μl of fresh growth medium was placed on top of them, and cultured at 37. The cells were decomposed and subjected to RNA analysis. HCV RNA transcript was determined by rt_QPCR in time. The content was adjusted to the cell gapdjj content. Results 20 (Fig. 4F) showed that the antiviral activity of about 95% L-peptide was lost in 1 hour at 37 ° C in 10% FBS. Under the same conditions, D-peptide can be completely stabilized for at least 4 hours. Thus, in addition to low immunogenicity and possibly oral bioavailability The peptide consisting of D-amino acid has the potential therapeutic advantage to improve serum stability. 115 200837075 50% inhibitory concentration (IC5G) and 50% fatality (LC50) IC50 of peptide 1 (concentration of peptide that inhibits HCV infection by 50%), greater than 95% pure peptide backup solution (3.6 mM in DMSO) diluted twice in DMSO - serially. Then obtained from one of the respective dilutions of the peptide Dilute 1:1 于 in Complete Growth 5 medium and mix in equal volumes of virus supernatant. The virus-peptide mixture was then used to infect Huh-7.5.1 cells (moi = 0.1) and cultured for 3 days at 37 °C. The percentage of HCV infection inhibition was determined by comparing the intracellular HCV RNA content in the virus-infected cells between the peptide-treated group and the DMSO control group. The cytotoxic activity of the peptide was determined by MTT cell toxicity assay according to the manufacturer's instructions (ATCC Cat #30-1010K), as described in the Methods section. The peptide concentration that reduces the OD toxin by 50% is indicated as LC50. As shown in Figure 4A, the L_isomer and D-isomer of peptide 1 are highly inhibited, showing IC5 〇 values of 〇·79μΜ and 0·32μΜ, respectively, with an LC50 value of about 100-300 times. high. The D-isomer is slightly reduced (ic5G), reflecting an increase in blood stability, because the antiviral activity of the L-isomer is reduced by about 2 logs after 1 hour of serum culture, and D- The isomer remains intact after at least 24 hours of culture (Fig. 4G). Peptide 1 also showed favorable toxicity profile data in vivo because L-isomers and D-isomers were administered at weekly intervals of up to 0.5 mg per 25 g of C57B1/6 mice. It was completely non-toxic when administered intravenously for 3 weeks. Importantly, peptide 1 was not immunogenic after repeated intravenous administration because antibodies against peptide 1 were not detected in the serum of these mice, as shown in Table 7 below. 116 200837075 0S ®§ as splash 091 sister nls strain 0S Bol^ eoot ms ms 00 s 3 0 8 r«

9·«ss JwstN rjn L<rz s 9·ε3 <F δ 0 (srs inch. SCNI 3 Oo-VO: CN: LLl 3 oo·inch Z: smss 3 δ 3 3τα s 3 2 2 r« s § Read § i 3 β 0 _ m tt*« te)s §s L<tz 0 « _tcirK: 卜.3⁄4 s ίβ β 0 S3 0 ss 3 3 0 s 3 2 s 3 in β 3 3 3 3 0.92 s 3 Sw s rs 3 P3i inch rnz 3 β i 8 0 s •3 sQow L%Z s 3 ss _ 0 00'« I inch·» s δ i loorsl s 言言_ Tls _ T 舯0_I _ . Tf Tf e# So T$Ql _ Ύ03 _ θ#σι. ws.. s$ nia —-a iTa mw,a 53 ii iw-i 13-Ί Ϊ3-Ί Is-T omaaspip^osp *^:_ςΌ 二诒wa *SSO 2 3 ά two w_aIf two ww-a 117 200837075 1 dimethylaminonaphthalene sulfonate · peptide 1 control dimethylamine thiol-PBS D · peptide 1 12921 0,118 HBV core control PBS core 05761 0.0386 «^ Peptide control HA pqs peptide 1.1305 0.0869 EV peptide peptide before harvesting peptide i rib 0314 0293 0251 0261 0.176 OJ80 0,107 0.108 peptide 1#2 ▲ 前胜% PBS 0.429 α437 0389 0394 025ί 0265 0.158 0.159 D·胜peptide1# 1 #jk前胜peptide1 PBS 0354 0345 0278 0271 0.159 (X163 0.104 0,108 DMSO before blood collection mmi PBS 0.526 0.497 0.401 0399 0285 0271 0.177 0.177 II si:i〇120 1:40 1:80 dimethylaminonaphthalene-flavor peptide-1 control dimethylaminonaphthalenesulfonyl-PBS D·peptide 1 L0206 0,12 HBV core control HBV s core PBS 0,566 0.0377 ΗΑ peptide control 0,7923 0.0506 D»peptide ί float 3 second blood collection D-peptide 1 ^ 0.411 0391 0269 0258 0,174 0.160 0.106 0.101 1> peptide 1 #2 second blood win Peptide 1 PBS 0278 0239 0^85 0272 0.197 0,190 0.116 0.120 peptide 1 #1·一^勒勒^血mu PBS 0.461 0.487 0374 0380 0229 0241 0.141 0.136 DMSO second blood sampling mki PBS 0.437 0.431 0374 0373 0211 0234 0J35 0.142 II s 1:10 1:20 1:40 1:80 Dimethylaminonaphthalenesulfonyl-peptide 1 control dimethylaminonaphthalene PBS PBS 1> peptide 1 1.0799 0.12 HBV core control HBV core PBS 0564 0.0381 HAJ1 peptide control ΗΑ peptide PBS 0.7923 0.0508 1> peptide 1 坍 second blood collection 1 > peptide 1 PBS 0377 (MX) 0304 0287 0201 0205 0.129 0.130 1> peptide 1#2 second blood collection D ·Peptide 1 PBS 0335 0292 0J07 0283 0210 0227 0.143 αΐ52 D·胜peptide1#1 Second blood collection D·* peptide 1 PBS 0.551 0.501 0 .426 0.407 0285 0291 0.182 0.184 DMSO Second blood collection peptide PBS 0411 0.414 0373 0361 0281 0279 0.171 0.176 Serum scale S § § ? §

118 200837075

谨 I would like to approve II dimethylamine __ PBS 1> peptide 1 1.0994 0.116 like 1 i 1 reading Qing core? BS 0.5815 0,0414 m PRC dirty PBS 0.9347 0.0784 S| |i §a ιάι PBS 0.351 0.385 0306 0.320 0.189 0.207 0.126 0J31 ;^: yT- f Vv) m〇91Γ0 16 ΙΌ 18ΙΌ 062*0 uro KrQ 893Ό Ship s| PBS 0.365 0,342 0.292 0.304 0,199 0.198 0,134 0.129 il a ui Less than 1ii pbs 0,302 0.274 0,259 0,247 0.177 0:182 0.114 0.118 li s 1:10 1:20 1:40 1:80 200837075 Example 7: Peptide toxicity Peptide cytotoxicity is based on the protocol provided by the ATCC MTT assay suite (Cat# 30-1010K) , determined by MTT cytotoxicity assay. Briefly, 5000-10,000 cells were seeded in each well of a 96-well plate. 5 After overnight growth, add 100 μl of fresh medium plus 20 μl of a 2-fold diluted peptide. The medium without the peptide was added to at least 3 wells as an untreated control. The cells were then incubated at 37 ° C, 5% CO 2 for 72 hours. After this incubation, 10 volumes of MTT solution (5 μg/ml in PBS) was added to each well and the cells were returned to the incubator. After 2 hours, the medium was removed, 10 150 μl of DMSO was added to dissolve the purple precipitate, and the wells were shaken at 150 rpm for 10 minutes. The background value of the 570 nm absorbance minus 670 nm is a reliable measure of cell death. The individual peptide cytotoxicity (LC5G) is defined as the peptide concentration that causes 50% cell death. The results (Fig. 5A) show that the LC5 values of the peptide 1 and the D_ form are substantially the same (excluding FBS of 3·8 μΜ 15 and 3·7 μΜ, respectively; and FBS containing 26·7 μΜ and 36, respectively). 8 μΜ Fresh human blood (treated with EDTA) was centrifuged 1000 g for 10 minutes to remove the supernatant and light yellow coating layer. Then the red blood cells were washed twice with PBS, with or without 16% FBS, and resuspended to a final concentration of 8%. A series of two-fold diluted peptides were prepared in 60 microliters of PBS on a 96-well microfluidic plate, and 60 microliters of human red blood cells in suspension with or without 20 FBS were added. The well plates were incubated at 37 ° C for 1 hour. After this incubation, 120 μl of PBS was added to each well, and the plate was centrifuged at 1 μg for 5 minutes. A whole 100 μl of the supernatant was transferred to a new 96-well micro-force plate. Microplate ELISA was used. The reader monitors the release of red blood cells by measuring the absorbance at 414 nm. In the wells, 0% and 120 200837075 100% hematocrit were measured in PBS and 0.1% Triton X-100, respectively. Calculated according to the following formula: 8 丨 4 legs of the peptide solution in the PBS of 8 4 丨 4 fine) / (〇 〇 ·ι% Cui Dun χ 〇〇 〇〇 A414nm - in PBS A4l4nm)] X 100. The results (Fig. 5B) indicate that the LCso values of the peptide and the d-5 peptide against human red blood cells are similar to each other when tested in the presence of Jinqing, and are similar to the LQo value of the hepatocyte cell line in the test tube. Importantly, the Lc5 〇 value of the anti-type cells is consistently 50 to 100 times higher than the EC5G value of each peptide. As for the preliminary measurement of cytotoxicity of peptides 1, 2 and 3 (refer to Table 4), a group of three mice (BALB/c mice, about 23 grams at 7 weeks old) were injected with 96 micrograms of 10 L type. Peptide 1 (approximately 4 mg/kg) was in 200 microliters of PBS (centrifuged for 3 minutes at 14,0 rpm before injection). In the control group, each of the three mice was given 2 μL of PBS containing 5% DMSO. The acute toxicity of the mice was monitored 3 hours after the injection. The results are summarized in the table below. Table 8: peptides 2 and 3 in C57BL/6 mice are non-toxic mouse weight (grams) (d.0) weight (grams) (d.3) weight (grams) (d.5) weight (grams (d.7) Weight (g) (d.10) DMSO-1 25.3 25.3 25.6 25.7 25.5 DMSO-2 23.1 24.4 24.6 24.8 25.1 DMSO-3 22.3 22.7 23.1 23.2 23.2 Peptide 1 22.2 22.3 22.8 23.1 23.5 Peptide 2 25.3 25.6 25.9 25.9 25.6 Peptide 3 24.1 24.1 24.7 24.7 24.7 No change in appearance, activity or behaviour observed. The mice were then weighed on days 0, 3, 5, 7 and 10. The weight gain of mice receiving peptide injection at an equal rate was used as a control group. 121 15 200837075 Example 8: Interaction between the physical properties of peptide 1 and its antiviral activity The secondary structure of peptide 1 (SEQ ID NO: 43) was used in cti.itc.virginia.edu/~cmg/Demo/wheel /wheelApp.html (last visit August 15, 2006) The spiral wheel application tool available online is divided into five parts. The resulting helical wheel (Fig. 6A) shows that peptide 1 is amphiphilic and has a water repellent surface and a hydrophilic surface. The secondary structure of peptide 1 was also analyzed using a circular two-color CD spectroscopy using an aviv model 62DS CD spectrometer (Avi Group, Inc., New Jersey Lake). The CD spectrum of the peptide was determined using a light-length test tube of 丨mm light path length at 1〇257. Three samples per sample were performed in 1 mM potassium phosphate buffer, ΡΗ 7.0, in the wavelength range from 1 nm to 260 nm. Data were collected at intervals of 60 nm/min at intervals of 〇·ΐ奈, expressed as mean molar ellipticity [q]. The peptide concentration was 50 μΜ. The characteristic spectrum of the south of the amphipathic α-helix was observed for the L form and the D form of the peptide 1 (Fig. 6). In addition, dimethylamino 15 naphthalenesulfonate can enhance the amphiphilic α-helical structure of peptide 1 (Fig. 6C). Thus, the peptide of the present invention may have a dimethylaminonaphthalene moiety covalently attached thereto. The secondary structure of each of the truncated derivatives of peptide 1 (Table 9) was analyzed using CD photolysis. The results indicate that deletion of two or four amino acids by the 丨c end of the peptide does not eliminate the α-helical structure of the peptide (Fig. 6). Conversely, the loss of two amino acids by the peptide 丄N and the booklet does eliminate the α-helix structure of the peptide (Fig. 6). The anti-HCV activity of the truncated variants of these succulents 1 was also determined. The results in the table below indicate that the antiviral activity of peptide 1 (L. form) has an interaction with its "_helix structure." 122 200837075 Table 9: C-terminal and N-terminal truncated derivative of peptide 1 peptide sequence SEQ ID NO : Anti-HCV Activity Peptide 1 (18-mer) SWLRDIWDWICEVLSDFK 43 + AC-16 Polymer SWLRDIWDWICEVLSD 94 + AC-14 Polymer SWLRDIWDWICEVL 92 + AC-13 Polymer SWLRDIWDWICEV 103 - AC-12 Polymer SWLRDIWDWICE 104 - ΛΟ10 SWLRDIWDWI 105 - AC-8 mer SWLRDIWD 106 - ΛΝ-16 mer LRDIWDWICEVLSDFK 107 - ΛΝ-14 mer DIWDWICEVLSDFK 108 - ΛΝ-12 mer WDWICEVLSDFK 109 - ΛΝ-10 mer WICEVLSDFK 110 - ΛΝ-8 mer CEVLSDFK 111 -

Example 9: Liposome-dye release assay analysis The peptides of Table 9 were also tested using the vesicle release assay 5 as discussed above. The results (Fig. 7) indicate that the antiviral activity of the various peptide 1 derivatives is related to the ability to cause release of the vesicle dye. Thus, the antiviral activity of peptide 1 is related to the release of the α-helical structure and the vesicle dye, and is summarized in the following table. 123 200837075 Table 10: Structure/Functional relationship of peptide 1 and its truncation C-terminal truncated peptide anti-HCV α-spiral dye release peptide 1 + + + AC-16 polymer + + + AC-14 + + + △C-12 mer - - - AC-10 mer - - - △C-8 mer - - - Ν-terminal truncated peptide anti-HCV α- Helical dye release peptide 1 + + + ΛΝ-16 mer - - - ΛΝ-14 聚 - - - ΑΝ-12 1 聚 - - - ΛΝ-10 聚 聚 - - - ΛΝ-8 聚 聚 - - -

Example 10: Structure of antiviral activity of succulent activity analysis of activity In order to determine whether the antiviral activity of peptide 1 is dependent on its primary amino acid sequence 5, four derivative peptides derived from peptide 1 are synthesized. To purity > 95%. The four peptides having the same amino acid composition include (1) peptide 1 sequence reversal (also referred to as anti-peptide); (2) water-repellent amino acid dispersal; (3) hydrophilic amino acid dispersal And (4) a derivative in which the aspartic acid residue (D) is replaced with a proline residue (P). The antiviral activity of the peptide is based on three peptide concentrations Μ8μΜ, 10 6μΜ and 2μΜ by HCV lesion reduction assay analysis, as explained above. The results are summarized in the table below, showing that the antiviral activity of peptide 1 has an interaction with the helical structure, but has no interaction with the primary amino acid sequence. 124 200837075 Table 11: Antiviral activity of the scrambled peptide 1 derivative. Sequence of peptides SEQ ID NO: Final concentration undiluted 1/3 dilution 1/9 SWLRDIWDWICEVLSDFK (La.a) 43 18μΜ 0±0 31±2 44 ± 8 KFDSLVECIWDWIDRLWS (La.a, reverse) 96 18μΜ 1±1 50 ±7 31±3 SWLRDIWDWICEVLSDFK (Da.a) ΝΑ 17μΜ 0±0 8±6 10±2 SIWRDWVDLICEFLSDWK (L-, water-repellent) 97 19μΜ 0±0 31±2 16±4 KWLCRIWSWISDVLDDFE (L-, hydrophilic dispersion) 98 20μΜ 0±0 8±6 15±6 SWLRPIWPWICEVLSDFK (L-? 2D/P) 91 19μΜ 9±3 21 ±8 41 ±2 False Sexual simulation ΝΑ 0.5 % DMSO 53 ±4 In other words, the HCV peptide library was synthesized by screening, and 13 peptides were identified to effectively inhibit HCV infection. For example, peptide 1 derived from the membrane-deficient 5 functional site of NS5A (NS5A-1975) is highly potent because a single dose of this peptide completely blocks HCV infection, EC5G = 289 nM, without cells Toxic evidence. At least 11 days after infection, the antiviral effect is significant. The peptide is most active when added to the cell along with the virus. The peptide is cultured in advance with the virus, which significantly reduces the attachment of the virus and reduces the infectivity of the virus, suggesting that the antiviral activity of 10 NS5A-1975 directly interacts with the virus and causes virus instability. The D-amino acid form of the peptide is completely active, and the D-form and L-form of the peptide are shown in the amphiphilic 〇:-helix structure in solution, which induces the penetration of human microlipids. A comparative analysis of a series of C-terminal and N-terminally truncated mutants (Sp. 3-13) against disease resistance, α-helicality and membrane decomposition activity (not shown) was performed. All peptides were synthesized to have a purity of > 95%, and their antiviral activity was compared by measuring their IC50 value 125 200837075. The helicity of the peptide was determined by circular two-color (CD) analysis using an Avi model 62DS circular two-color (CD) spectrometer (Ivey Group, Lake N.). The spectra were measured at 25 ° C using a 1 mm optical path length optical test tube. For each sample in the range of 19 〇 nanometer to 26 〇 nanometer wavelength range: underscan in 1 mM mM phosphate buffer with or without 5〇% trifluoroethanol (TFE) . The data rate of the data is (9) nm/min at 〇 ι 不 间隔 interval collection 'in terms of average molar ellipticity (7). The percent peptide concentration of the peptide is as calculated by Chen et al., Biochemistry 13, 3350, 974). The results are shown in the table below. 10 126 200837075

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40.7 ND 38.1 37.0 50.0 302 42.4 39.3 42.0 36.7 3L4 35.7 31.9 0.85 0.48 0.50 0.40 Η Η § SK Η 13.5 12.5 13.0 L-Anti-D-Anti-Hydrophilic Dispersion Dispersion S i IS fiss ^ ® ® ^sts mT\ rFT&gt o5T\S shou shoulder shoulder vg §! circle, uuo Ills _ IIIL Hill illusion "泠1 1 stggg 1 gs Q θ 1 α Θ g IIi|II lllll Mil r5 ^ S rn r 〇 § § ggg Q θ Θ III _ III 00 ζ /3 ζ /3 Sg 128 200837075 16-mer SWLRDIWDWICEVLSD (SEQ ID NO: 94) (peptide 3 of Table 12) retains intact antiviral activity, α-helical properties and membrane breakdown properties (Table 12b). The antiviral activity of the caudate truncated and C-terminal truncated peptides has an interaction with its membrane permeation activity and the amphipathic α-helical structure. The truncated mutant lost all three activities at the same time, suggesting that it may be functionally linked to the activity of the peptide to inhibit HCV infection. As shown in the spiral diagrams of Figures 6 and 6 , peptide 1 has strong amphiphilicity. In order to determine whether amphipathicity is required for its antiviral activity, the inventors tested two by exchanging the position of the amino acid (underlined position in Table 12c) while retaining its amine 10-acid composition and alpha-helicality. Two variants of reduced affinity (peptides 18 and 19). As shown in Table 12c, the antiviral activity of these peptides was reduced by a factor of 5 to 30. These results are contrary to the antiviral activity of the mutant strain, and the water-repellent amino acid or hydrophilic amino acid is selectively broken up while retaining its amphiphilicity (Table 12d, peptides 25 and 26). Collectively, these results suggest that 15 amphipathic is required for antiviral activity. But the deficiency is due to three amphipathic α-helical peptides that have previously been clearly characterized, such as Ma Jianing 2 (Bechinger et al., Protein Science 2, 2077-84 (1993)), and Apolipoprotein 18A (Chmig et al. , J Biol Chem 260, 10256-62 (1985)) and the peptide 1 analog from BVDV (Sapay et al., Biochemistry 45, 2221-33 2〇 (2006)) does not have anti-HCV even at high concentrations. Antiviral activity (Table 12c, peptides 20-22). In order to determine whether the primary amino acid sequence is required for antiviral activity, a series of peptides having the same amino acid composition but different primary sequence as the parent peptide were tested. As shown in Table 12d, the peptide containing the reverse sequence (anti-) (peptide 23 and 129 200837075 24) or the broken sequence (peptides 25 and 26) but retaining the amphipathic and α-helix degree is similar. The substance is completely active. These results indicate that the antiviral activity of the peptide is independent of its primary amino acid sequence, as long as the amphipathic helical structure is maintained. 5 In order to determine whether the antiviral activity is strictly dependent on the amino acid composition, the inventors compared the activity of several peptides and peptide analogues from the six HCV genotypes, and the amino acid sequence and composition were not strictly retained. Sex. As shown in Table 12e, the antiviral activity of the peptide was found to vary within a 10-fold range, with genotypes la and 3a (peptides 28 and 29) having the highest activity, while genotypes 4a and 5a (peptides 30 10 and 31). ) has the lowest activity. These results suggest that the amino acid composition of the peptide apparently affects its antiviral activity. It is noted that the cysteine residue in peptide 1 may result in the formation of a disulfide bond between the peptide monomers, which are required for antiviral activity (Table 12f). Example 11: Effect of peptide on VSV infection 15 To determine whether the antiviral activity of peptide #1 is specific for HCV, a similar experiment was performed on other enveloped viruses such as vesicular stomatitis virus (V S V). Two assay assays were used to test the antiviral activity of peptide #1 against vsv. Blocking of infection: To test whether peptide 1 blocked vsv infection, peptide 18 at a final concentration of 18 μM and VSV at 1 to 10,000 pfu/ml were simultaneously added to Huh-7 cells. Paratope peptides and HCV (10,000 pfW ml) were added to the cells as controls. After 4 hours of adsorption at 37 ° C, the virus-peptide inoculum was removed. The cells were washed twice, and 120 μl of fresh growth medium was plated and cultured at 37 ° C for 3 days. VSV infection and HCV infection were assessed by immunocytochemistry using viral cytopathic effect 130 200837075 (CPE) and antibodies against HCV E2 protein, respectively. Virucidal activity: To determine whether peptide 1 has antiviral activity against VSV, peptide 1 was diluted to a final concentration of 18 μM in complete 5 growth medium containing 2 X 105 pfu (ffu) per milliliter of VSV or HCV. The virus-peptide mixture was then incubated at 37 ° C for 4 hours. The VSV and HCV viral valences were then determined by a series of dilutions, assessed by viral cytopathic effect (CPE) and immunostaining using anti-HCV E2 protein antibodies. The results (Fig. 8) indicate that peptide 1 did not block VSV infection and had virucidal activity against VSV. Further experiments indicated that peptide 1 did not block infection by pandemic influenza virus, vaccinia virus, Borna disease virus, lymphocytic choriomeningitis virus or adenovirus (data not shown). Example 12: Effect of peptide on dengue-2 infection The following experiment was conducted to determine which peptide could inhibit dengue-2 virus infection. The peptides tested are shown in the table below. 131 200837075 Table 13 Sequence of peptide name SEQ ID NO: 2013 peptide SWLRDIWDWICEVL 92 2015 peptide SWLRDIWDWI 105 2054 peptide SWLRDIWDWICEV 103 2017 peptide LRDIWDWICEVLSDFK 107 2018 peptide DIWDWICEVLSDFK 108 L-2022 (L-7208) peptide SWLRDIWDWICEVLSDFK 43 D -2022 (D-7208) peptide SWLRDIWDWICEVLSDFK NA L-7208 (2D — 2 Pro) SWLRPIWPWICEVLSDFK 91 L-7208 HS SIWRDWVDLICEFLSDWK 97 3222 SWLRDIWDWISEVLSDFK 127 3226 SWLDRIWRWICKVLSRFE 128 3227 SWLDDIWDWICEVLSDFE 129 3228 SWLRRIWRWICKVLSRFK 130 3229 SWRLDIWDWICESVLDFK 119 3310 DIWDWICEV 121 3311 RDIWDWICEV 122 3244 DWLKAFYDKVAEKLKEAF 120 6938 LYGNEGCGWAGWLLSPRG 6 Enzyme-linked immunosorbent assay: Vero cells (80,000 cells/well/ml) were seeded in 24-well plates 24 hours prior to infection. Cell lines were exposed to dengue-2 in the presence of increasing concentrations of 5 peptides (or DMSO as a control)

Vero cells. Virus and peptide were not removed during the entire culture period (cells were not washed). The infection was analyzed by ELISA 5 days later, and the number of dengue-2 shells released from the supernatant of infected Vero cells was measured by ELISA to analyze the infection. 132 200837075 Fluorescent lesion assay: Vero cells were seeded in % well plates 24 hours prior to infection. Cells were exposed to dengue-2 in the presence of increasing concentrations of peptide (or DMSO as a control). The virus and peptide were washed off 2 hours after infection. Supernatants were collected every 3 感染 after infection and added to fresh Ver〇 cells for fluorogenic lesion analysis. Newly infected Vero cells were mixed with 4% formaldehyde after 3 days. Then the cells are dengue

The Env antibody was stained and then stained with a secondary antibody conjugated with an Ayxa-flu〇r dye. The number of lesions was calculated using a fluorescence microscope. The results are summarized in the table below and in Figure 9. Table 14: Inhibition of dengue infection by ELISA peptide sequence a-DEN ENV a-shell 9A7 2022 peptide (20 μΜ) SWLRDIWDWICEVLSDFK (SEQ ID NO: 43) 97.8 98.02 2022 peptide (5 μΜ) 28.0 50.02 2022 win Peptide (1.25 μΜ) 0 0 2013 peptide (20 μΜ) SWLRDIWDWICEVL (SEQ ID NO: 92) 97.8 98.3 2013 peptide (5 μΜ) 29.65 22.7 2013 peptide (1.25 μΜ) 0 11.4 2017 peptide (20 μΜ) LRDIWDWICEVLSDFK ( SEQ ID NO: 107) 74.83 82.2 2017 peptide (5μΜ) 33.64 16.01 2017 peptide (1.25 μΜ) 10.24 12.82

As shown in Tables 14 and 9, dengue infection was inhibited by peptides present in a dose-dependent manner. Approximately 1% of dengue virus infection inhibition was observed at a concentration of 20 μM (Fig. 9). Intracellular FACS assay: Vero cells were seeded at 133 10 200837075 6-well plates 24 hours prior to infection. The cells were exposed to dengue-2 in increasing concentrations of peptide (or dms® as a control). The virus and peptide were washed off 2 hours after infection. After infection, 3 cells were taken for intracellular staining. Cells were stained with appropriate isogenic control, dengue, dengue, or tubulin antibodies. Cells were analyzed by FACS. 5 The results obtained using the concentration of 2〇μΜ peptide are shown in Table 15. The results of 1.25 μΜ to 20 μΜ are summarized in the line graph shown in the ioa-B diagram. Figure 10B-D shows that the potent inhibitory effect of dengue virus infection is related to the amphipathic structure of the peptide structure, rather than to the exact amino acid sequence of the peptide. Thus, the following peptides have a high inhibitory effect on dengue virus infection: peptide 1 (also known as 10 2022 and L-7208, SEQ ID NO: 43), peptide 3222 (SEQ ID NO: 127), peptide 3226 (SEQ ID NO: 128), peptide 3228 (SEQ ID NO: 130), peptide L-7208 2D to 2 Pro (SEQ ID NO: 91) and L-7208 HS have a disintegrated water-repellent amino acid ( SEQ ID NO: 97). Table 15: Detection of dengue infection by FACS 15 *PI-based Propidiumiodide staining for detection of dead cells; n/A- not applicable for % of intracellular FACS staining from 3 cells after infection GATED % dead cell sequence SEQ ID NO: isotype control dengue α-shell (9Α7)) a-tubulin PI* virus-free control N/AN/A 1.29 0.08 91.96 1.00 DMSO N/AN/A 0.39 41.26 96.67 1.00 2015 peptide SWLRDIWDWI 105 0.85 37.21 97.30 1.2 2054 peptide SWLRDIWDWICEV 103 1.68 2.61 97.35 3.04 2018 peptide DIWDWICEVLSDFK 108 0.97 12.07 96.83 0.76 L-2022 peptide SWLRDIWDWICEVLSDFK 43 0.68 0.82 95.65 L88 D-2022 peptide SWLRDIWDWICEVLSDFK N/A 0.77 1.35 92.60 3.83 6938 peptide LYGNEGCGWAGWLLSPRG 6 0,69 42.41 96.32 4.05 134 200837075 As shown in Table 15 and Figure 10A-B, dengue infection is inhibited by the presence of peptides in a dose-dependent manner. Approximately 100% inhibition of dengue virus infection was observed at a concentration of 20 μM (Fig. 10A-B). Fluorescent lesion assay: Vero cells were seeded in 5 96-well plates 24 hours prior to infection. Cells were exposed to dengue-2 in the presence of increasing concentrations of peptide (or DMSO as a control). The virus and peptide were washed off 2 hours after infection. Supernatants were collected every 3 days after infection and added to fresh vero cells for fluorescence lesion analysis. After 3 days, the newly infected Vero cells were mixed with 4% A. The cells were then stained with antibodies against dengue envelope proteins, followed by staining with an arsenal fluorescent dye-conjugated secondary antibody. The number of lesions was calculated using a fluorescence microscope. The results (not shown) confirmed that the peptide strongly inhibited dengue virus infection. At a concentration of 2 〇μΜ, approximately 1% inhibition of dengue virus infection was observed. Example 13: peptide 1 has potent antiviral activity against West Nile virus infection. In this study, test peptide 1 and peptide 2012 (SEQ ID NO: 94) 15 anti-West Nile virus (WNV) belongs to a flavivirus ( Flavivirus) activity. A549 cells were infected with 102 to 10 PFU/ml WNV (New York germline) in the presence of 0.5% DMSO, peptide 1 or peptide 2012, where the peptide final concentration was 18 μM in 0.5% DMSO. After 3 weeks of infection at 37 ° C, the cells were fixed and subjected to immunoperoxidase staining to detect WNV protein. Results (Fig. η) 20 shows that the cell monolayer with 1 〇 5 pfu/ml treated with DMSO was almost completely destroyed, and all cells in the lower valency well exhibited WNV protein. Conversely, the monolayer in the peptide-treated cells is intact with little or no detectable WNV protein. In particular, the WNV protein staining intensity was the same as the unstained negative control well, regardless of the viral inoculum dose. Such a knot 135 200837075 The authenticity peptide 1 (SEQ ID NO: 43) and the peptide 2012 (SEQ ID NO: 94) have potent antiviral activity against WNV infection. Example 14: Streptavidin 1 and its variant strain inhibit HIV infection This example shows that the peptide of the present invention inhibits infection by human immunodeficiency virus. Materials and Methods Virus was generated by transfer of the nucleoside vector 293T cells using Genejuice (Novagen). The virus supernatant was harvested 48 hours after infection and filtered through a 0.2-μΜ pore size filter to remove fine debris. Viral inoculum was standardized by p24 (HIV-l capsid) enzyme-linked immunosorbent assay (PerkinElmer Life Sciences). The TZM-bl reporter cell is a CD4+CXCR4+CCR5+HeLa cell containing the LacZ gene driven by the HIV-1 LTR (starter gene). At the time of infection, HIV-1 Tat protein was produced and HIV-1 LTR was activated. TZM-bl cells (80,000 cells/well/ml) were seeded in 24-well plates 24 hours prior to infection. The following peptides were tested for HIV infection inhibition. 136 200837075 Table 16: Sequence of peptide peptides tested for anti-HIV activity SEQ ID NO: 6938 LYGNEGCGWAGWLLSPRG 6 2015 SWLRDIWDWI 105 2054 SWLRDIWDWICEV 103 2018 DIWDWICEVLSDFK 108 L-2022 (L-7208) SWLRDIWDWICEVLSDFK 43 D-2022 (D-7208) SWLRDIWDWICEVLSDFK ΝΑ L-7208 HS SIWRDWVDLICEFLSDWK 97 L-7208 (2D — 2 Pro) SWLRPIWPWICEVLSDFK 91 3222 SWLRDIWDWISEVLSDFK 127 3226 SWLDRIWRWICKVLSRFE 128 3227 SWLDDIWDWICEVLSDFE 129 3228 SWLRRIWRWICKVLSRFK 130 3229 SWRLDIWDWICESVLDFK 119 3244 DWLKAFYDKVAEKLKEAF 120 3310 DIWDW1CEV 121 3311 RDIWDWICEV 122

Note that the L_2022 peptide (also referred to as "peptide 1" and L-7208) has the same sequence as the D-2022 peptide (also known as D-7208), but the L-2022 peptide system 5 is composed of L. amine. The acid consists of a D-2022 peptide consisting of a D-amino acid. Further, L-7208 HS peptide (SEQ ID NO: 97) and 3229 peptide (SEQ ID NO.. 119) have the same amino acid composition as L-7208 peptide (SEQ ID NO: 43). However, although the L-7208 HS peptide has an amphiphilic structure, the 3229 peptide is not. The 3310 peptide (SEQ ID NO: 121) and the 3311 peptide (SEQ ID NO: 10 122) contain the central 137 of the highly active peptide L-2022 (SEQ ID NO: 43), respectively. 200837075 Nine and ten amino acids Sequence, but too short and therefore very low activity. The 3244 peptide and the 6938 peptide were used as controls. The cells were exposed to twist in the presence of increasing concentrations of peptide, and DMSO without peptide was used as a control. The peptide is added to the virus and then 5 is added to the target cells. Although the preliminary study utilized the HIV branching system B CCR5 virus (JR-CSF), the experimental lines described below were performed with other HIV strains, including NL4-3 (derived from R9BaL, ADA or YU2 (all showing CCR5 tropism). CXCR4) Recombinant HIV germline of the backbone protein and envelope protein (GP120). 10 Virus and peptide were washed off 2 hours after infection. Infected cells were analyzed by analysis of /3 -galactosidase activity 48 hours after infection. For galactosidase activity, infected TZM_bl cells were washed twice with 1 ml of phosphate buffered saline and dissolved in 100 mM potassium phosphate containing 0.2% Triton X-100, pH 7.8. The plates were stored at -80 ° C for 16 hours, thawed on ice, and 20 μl of the lysate was transferred to a 96-well plate to detect/5-galactosidase activity. Galanton-Star Enzyme Matrix (Applied Biosystems, Bedford, MA) in reaction buffer diluent (1 mM mM sodium phosphate pH 7.5, 1 mM MgCl2, 5% sapphire (Sapphife) A reaction buffer was prepared by diluting 1:50 in a IITM accelerator. One hundred microliters of reaction buffer was added to 20 microliters of the lysate, and after 30 minutes, the luminescence was measured in a microplate luminometer for a period of one second. Results The peptide of the present invention is a potent inhibitor of HIV infection. As shown in Figure 12, the 2054, 2018, L-2022, and D-2022 peptides having SEQ ID NOS: 103, 108, and 43 are substantially 100% inhibited when present at a concentration of 20 micromolar. infection. Even lower concentrations of 2018, L-2022 and D-2022 (SEQ ID NO. 108 and 43) are also active in the south. In particular, both isoforms of 2Q18 peptide and 2022 peptide inhibited 9〇-1〇〇% HIV infection at 5 micromolar concentrations. 2022 (SEQ ID NO: 43) The D-isomer of the peptide was maintained at a low concentration; 5 1.25 micromolar concentration maintained high activity (Fig. 12). • Figure 13 further shows the efficacy of this peptide. In particular, Figure 13 shows

• At a concentration of 20 micromolar, the peptide l-7208 HS • : (SIWRDWVDLICEFLSDWK, SEQ ID NO: 97) is equivalent to the highly active SEQ ID NO: 43 peptide. L-7208 HS (SEQ ID 10 NO · 97) has the same amino acid composition as the highly active SEQ ID NO: 43 L-2022 peptide, but the water-reducing amino acid in the L-7208 HS peptide has been After being broken up, although it retains amphipathic, it has a different sequence than the SEQ id NO: 43 L-2022 peptide. These data indicate that although the two fraternities of the peptide are important for the activity, the exact sequence of the peptide is not particularly limited. 15 Use HIV branching system B CCR5 virus (JR-CSF) and NL4-3 ^ (CXCR4) backbone protein and envelope protein (GP120) derived from _ · R9BaL, ADA or YU2 (all showing CCR5 tropism) Recombinant HIV germline observations • The same results. In addition, the peptide is highly effective in inhibiting different types of cell infection. Inhibition of HIV R9BaL infection in 293 T cells is shown in Figure 13A, and similar results were obtained for HIV R9BaL infection of 20 CEM T cells as shown in Figure 13B. The peptide destroys HIV-1 and releases the viral contents to the culture medium. Thus, Figure 14A shows that after treatment of HRM with peptide L-7208 (SEQ ID NO. 43), a large number of sputum shells are released from the virions, suggesting that the peptide is available at 139 200837075

Extracellular destruction of viral particles. In contrast, very little or no release of the HIV-1 shell (Fig. 14A) after treatment with DMSO or with peptide 6938 (SEQ ID NO: 6) was used as a negative control. Matching this observation, using the peptide L-7208 (SEQ ID NO: 43), after comparison with DMSO or treatment with peptide 6938 (SEQ ID NO: 6), the virus-associated HIV-1 shell was observed. Significantly reduced (Figure 14B). Finally, Figure 14C shows that the HIV-1 capsid is internalized inside the cell after treatment with virions using DMSO and 5 or 10 micromolar peptide 6938 (SEQ ID NO: 6) or L_7208 (SEQ ID NO: 43). The percentage. When the viral particle 10 was treated with 5 or 10 micromolar peptide 6938 (SEQ ID NO: 6) prior to infection, approximately 100% of the control amount of the HIV-1 shell was internalized, but the peptide L- was used. Treatment with 7208 (SEQ ID NO: 43), the internalization of the HIV-1 shell was inhibited by up to 10-fold. Figure 15 shows that the peptide having this amphiphilic structure similar to the amphiphilic structure of the peptide L-7208 (SEQ ID NO: 43) can also strongly inhibit HIV infection. Such amphiphilic peptides having potent anti-HIV activity include peptide 3222 (SEQ ID NO: 127), peptide 3226 (SEQ ID NO: 128), peptide 3228 (SEQ ID NO: 130), peptide L- 7208 2D to 2 Pro (SEQ ID NO: 91) and L-7208 HS 20 (KWLCRIWSWISDVLDDFE, SEQ ID NO: 98) with a hydrophilic amino acid fragmented. Therefore, as observed for HCV and for dengue virus, the antiviral activity of the peptide against HIV is determined by the amphipathic nature of the peptide. In addition, when HIV infectivity is inhibited, the HIV shell is released into the supernatant, indicating that the peptide destroys or dissolves the HI V virions. 140 200837075 Therefore, these peptides derived from the hepatitis C virus genome are highly potent anti-HIV agents that facilitate the suppression of HIV infection. Example 15: Antiviral specificity and range of peptide 1 In order to determine whether the antiviral activity of the peptide 1 is HCV specific, the antiviral activity of a large series of other viruses listed in Table 17 is as follows. Various concentrations of peptide or DMSO were added to a virus stock with a predetermined infectious force (MO5 ffl^TCID5〇/ml) using a 2-fold serial dilution starting at 18-20 μM. The virus-peptide mixture and the virus-DMSO mixture were incubated at 37 ° C for at least 1 hour prior to addition to sensitive cells, unless otherwise specified. Parallel 10, peptide and HCV (10,000 ffu/ml) were added to Huh-7.5.1 cells as a positive control for antiviral activity. After 2-4 days of infection, cultures were assessed by detecting cell pathogenic effects (CPE) or by antibody immunostaining against the corresponding viral proteins, as described in the Methods section. The HCV infection is characterized by JFH-1 (genotype 2a) and a structural region containing the isolated isolates of genotypes 15 la (H77), lb (coni) and additional genotype 2a pure strains (J6CF). Chimeric virus is carried out. To make chimeric HCV genomes, a recombinant PCR approach was used. Such as

Pietschmann et al., Proc Natl Acad Sci USA 103, 7408-13 (2006), replacing the corresponding JFH-1 core-NS2 with pUC-vJFH via the corresponding sequence 20 from J6CF-, H77 and Coni The region produces J6CF-, H77 and Conl-JFH-1 chimeric HCV genomes. Infectious JFH-1 virus and chimeric virus were produced by transferring the in vivo HCV RNA synthesized into the tube to Huh-7.5.1 cells; the virus preparation containing i〇4_105ffu/ml is such as Zhong et al., Pr〇c Prepared as described in Natl Acad Sci USA 102, 9294-9 141 200837075 (2005). Since HBV was not contagious in the test tube, the inventors used quantitative ELISA to examine the effect of the peptide on the antigenicity of HBV, and as described above, the effect of the peptide on the HBV DNA content was examined by quantitative PCR analysis. 5 for dengue-2 infection, courtesy of Christopher Aiken, Tennessee

Vero cells from the Vulcan Vanderbilt University School of Medicine were seeded in 6-well plates 24 hours prior to infection. Cells were cultured in the presence of dengue-2 (provided by Richard Kinney, Atlanta, Georgia, CDC, Richard Kinney) in increasing concentrations of peptide (DMSO as a control). The virus and peptide were washed off 4 hours after infection. After infection, 10 cells were harvested for intracellular FACS staining. Cells were stained with the appropriate isogenic control group (BD Biotech, Pharmingen, California La Jolla), or with dengue-specific antibodies (Chemicon, Temecula, CA) ) staining, analysis by intracellular FACS assay (FAC FACSort, BD Biotech, San Jose, CA). Intracellular FACS assay of dengue-2 virus 15 Analysis (IFSA) was performed by S. Selvarajah and P. Gallay, and the data were not published. For HIV-1 infection, use wild-type pNL4.3 provided by Malcolm Martin through the NIH AIDS Research and Reference Reagent Program or an infectious molecular strain derived from PNL4.3, which has been BAL and JR- Encapsulated gene replacement of CSF (Bobardt et al, J Virol 81, 295-405 20 (2007)). Molecular pure strains were generated using genetic genomics (Novagin, San Diego, Calif.) to produce infectious HIV by transfer of liposome-mediated 293 T cells. The TZM-bl reporter cell is a CD4+CXCR4+CCR5+HeLa cell containing the LacZ gene driven by the HIV-1 LTR promoter gene (by John C. Kappes, Xiaoyun Wu, and Tranzyme Inc 142 200837075). Available through the NIH AIDS Research and Reference Reagent Program). The cells were exposed to H i i in the presence of increasing concentrations of peptide (D M S Ο as a control). The virus and peptide were washed off 2 hours after infection. Infected cells were analyzed by galactosease 5 activity 48 hours after infection as described by Chatterji et al., j Bi〇i chem 280, 40293-300 (2005). For measles infection, the measles virus wild-type germline (WTF) is grown on BJAB cells and is used to infect Vero cells that express human SLAM-receptors in the presence or absence of peptide or DMSO with moi 0.3. All reagents were supplied by T.B.H. Geijtenbeek, VU University Medical Center, Amsterdam, The Netherlands. At 48 hours post-infection, WTF infection was determined by staining cells with anti-Μν η antibodies (CV1, CV4) (Kemicon, California, Temecula), followed by Witte et al., J Virol 80, 3477- Cells were stained with goat anti-mouse IgG-FITC antibody as described in 86 (2006). For West Nile virus infection, a series of 2-fold diluted peptides 1 was added to 15 dilutions of lxlO5 pfu of WNV-NY stock. The virus peptide mixture was added to Huh-7 cells at moi = 1 and cultured at 37 ° C for 48 hours. As described by Fredericksen et al., J Virol 80, 2913-23 (2006), immunostaining with anti-WNV polyclonal antibodies allows the infected cells to become visually visible. Other studied viruses include HSV-1 germline F (ATCC 20 Cat#VR-733), bovine viral sputum virus germline NADL (VR-534), human crown virus germline 229E (VR-740), human kesha F5 gene Faulkner (VR-185), human respiratory fusion virus line Long (VR-26), human rhinovirus 14 lines 1059 (VR-284), human rotavirus line WISC2 (VR-2417) And the vesicular stomatitis virus strain Indiana Lab 143 200837075 (VR-1238). These viruses were purchased from the American Type Culture Collection and used Huh-7 cells as recommended by the supplier or as permitted by the cells, in Huh-7, MDCK (ATCC Cat#CCL-34), MRC- Cultured in 5 (CCL-171), HeLa (CCL-2), Hep-2 (CCL-23), HeLa, ΜΑ-104 5 (CRL-2378), and Huh-7 cells. Can be expressed by CMV promoter

Recombinant adenoviral subtype 5 of driven GFP is supplied by U. Protzer, Coron University, Sprinzl et al, J Virol 75, 5108-18 (2001), and is used to infect human HEK293 cells (CRL-1573). The recombinant vaccinia virus line was supplied by B. Moss, ΝΙΑΠ 3 (Thimme et al, J Virol 77, 68-76 (2003)), and 10 was used to infect Huh-7 cells. The mouse-adapted influenza A/WSN/33 line was provided by Adolfo Garcia-Sastre (Falcon et al, J Gen Virol 86, 2817-21 (2005) (Sinai Hill Medical School, New York, NY)). LCMV (Armstrong) and BDV (He80) and Vero cell lines are provided by Juan Carlos de la Torre (cf. Clemente et al, J Virol 81, 5968-77 (2007) 15 and Buchmeier et al, Virology 113, 73-85 ( 1981)) (Immunary Department, TSRI 'La Jolla, California). All three viruses were used to infect Vero cells. To determine if the selected peptide inhibits viral infection, a series of 2-fold diluted peptides (18-20 μM) or DMSO is added to the virus preparation with the predetermined infectivity (1-105 ffu or TCKW ml) at 37 °C. Incubate for 1 hour, then add 20 to the susceptible cells for 24 days, at which time the comparative assessment of cell pathogenic effects (CPE) (BVDV, Scravus virus, HSV1, crown virus, influenza virus, rhinovirus) , rotavirus, RSV, vaccinia virus, and VSV) or immunostaining using anti-corresponding viral protein antibodies [Bona virus (Ref. Buchmeier et al., Virology 113, 73_85 (1981)), LCMV 144 200837075 ( Each culture was evaluated separately with reference to Clemente et al, J Virol 81, 5968-77 (2007)), adenovirus (see Sprinzl et al, J Virol 75, 5108_18 (2001)). The results indicated that at a concentration of 18 μΜ, peptide 1 against adenovirus 5, Bona disease 5 virus, bovine viral sputum virus, crown virus, krusa virus, herpes simplex virus 1, influenza virus, lymphocyte Choroidal plexus meningitis virus, rhinovirus, rotavirus, vaccinia virus or vesicular stomatitis virus had no significant effect; or had no significant effect on the antigenicity and DNA content of hepatitis B virus. 0 Conversely, peptides strongly inhibit the infectivity of other members of the Flaviviridae family, including West Nile virus and dengue-2 virus. Unexpectedly, peptide 1 strongly inhibits the infectivity of paramyxovirus, measles virus and respiratory syncytosis virus and human immunodeficiency virus. The ICso values of peptide 1 for these viruses are shown in Table 17, below. 145 200837075 Table 17 Antiviral specificity and effective range of viricides Viral envelope gene Ι〇5〇(μΜ) HCV(JFH-1) genotype 2a is RNA+ 0.6 HCV (H77 envelope) genotype la is RNA+ 3.9 HCV (Coni envelope) genotype lb is RNA+ 1.6 HCV (J6CF envelope) genotype 2a is RNA+ 1.1 dengue virus is RNA+ 2.0 West Nile virus is RNA+ 4.5 Alpha virus is RNA+ 2.7 Respiratory fusion virus is RNA-4.5 human immunodeficiency virus Is RNA+ 1.3 Adenovirus No DNA >18 Bona disease virus is RNA- >18 Bovine viral sputum virus is RNA+ >10 Crown virus 229E is RNA+ >18 克沙奇病毒NORNA+ >18 Hepatitis B The virus is DNA >18 Herpes simplex virus 1 is DNA >10 Influenza virus is. RNA- >18 Lymphocytic choriomeningitis virus is RNA- >18 Rhinovirus No RNA+ >18 Rotavirus WISC2 No RNA+ >18 Vaccinia virus is DNA >18 Vesicular stomatitis virus is RNA- >18 Example 16: peptide 1 has strong antiviral activity against HIV test peptide 1 D-isomer with a group of hosts It also has two parents The ability of the viral peptide to block HIV infection. Method 146 200837075 Cells • Unheated DC lines were cultured as described by Geijtenbeek et al., Cell 1 〇〇, 587-97 (2000). To put it bluntly, by using the Fic〇11 gradient and subsequently using the MACS system (Mitan Yisheng Technology Co., Ltd. (New Zealand 11001 GGmbH), Germany Baiji S. Grabeck) CD14 selection step, shallow The yellow 5 coating separates human blood mononuclear cells. Purified monocytes at 500 units/ml of interleukin-4 (IL-4) and 800 units/ml of granulocyte-macrophage colony stimulating factor (GM-CSF) (Schering-Plough, In Brussels, Belgium, the differentiation into an immature DC. On the sixth day, flow cytometry was used to verify the 10 phenotype of cultured DCs. DCs exhibit high MHC classes I and II, CDllb, CDllc, ICAM-1 and low CD80, CD83 and CD86. PBMC was isolated from a light yellow coating and activated using phytohemagglutinin (3 μg/ml). At the third stage, the cells were washed and co-cultured with IL_2 (1 unit/ml). The macrophage cell line derived from CD4+孓 lymphocytes and monocytes is as described above for separation 15 and determining feature 17. TZM_bl cells can express CD4, CXCR4 and CCR5, and contain the integrated lacZ gene driven by HIV-1 LTR (Wei et al., Antimicrobial Agent Chemotherapy 46, 1896-1905 (2002)). A genital epithelial cell (PGEC) is provided by B. Kahn of the Scripps Clinic Women's Department. Rotate the swab swab towards the vaginal wall and collect millions of cells for each individual. The cells were placed in sterile PBS for 20 minutes, maintained at 4 ° C, and sent to the laboratory. After centrifugation (3 〇〇g for 5 minutes), the cell pellets were contained in 丨mg/ml liter (Sigma) and 〇15 mg/ml N_α_p-phenylsulfonyl _L_ionide Acid chloromercapone (Sigma) 1 gram / soar collagenase _ dispase (R0 Che Molecular Biochemicals) in it digestion 147 200837075 1 hour. The digest was centrifuged (1,000 g for 20 minutes) and resuspended in 250 mg/ml PBS/BSA. After additional centrifugation, the pellet was resuspended in 5 mg/ml PBS/BSA and loaded on a 50% Percoll gradient. Then, as described previously in Bobardt et al, J Virol 81, 395-405 (2007), PGEC was isolated from the FACS stained fine sputum classification, containing 10% FCS and epithelial cell growth supplement (1) DMEM F12 medium (Sigma) was propagated into a collagen type I-coated T-25 flask. The subculture of PGEC was used less than three times before use to maintain its original characteristics. : 293T cells were infected with protoplasmic plastids (9 μg) and VSV-G by membranous 10 (1 μg). On day 2, The VSV-G pseudotype virus was used to acutely infect Jupiter cells. After infection, the virus was harvested and the p24 content was determined by ELISA (Berkin Emma Life Sciences). The following proviral constructs were used: wild type pNL4. 3 (X4); pNL4.3-BaL (R5) in which wild-type NL4.3 is membrane-exchanged with R5 BaL envelope; pNL4.3-^Env lacks gpl60; 15 pNL4.3-eGFP (X4); and pNL4.3- BaL-eGFP (R5) encodes the GFP gene instead of the Nef gene. One HIV-1, HIV-1 drug resistance, HIV-2, SFV and SHIV virus lines are obtained through the NIH AIDS Research and Reference Reagent Program, and are PHA/ Amplification in IL-2 stimulated PBMC. Infection · TZM cells (1〇〇, 〇〇〇 cells / 20 ml) were exposed to HIV (1 Nike p24) for 4 hours in the presence or absence of peptide 1 Washing, infection was measured by/5-galactosidase activity 48 hours after infection. No cytotoxicity was observed in >1〇〇μΜ peptide 1 without washing (data not shown). DC, CD4+ Τ-lymph Cells or giant scorpion cells (0·1 X 106 cells) were exposed to virus (1 ng p24), washed three times with medium, and cultured in flat-bottom 96-well plates. After different days, 148 were collected. 200837075 Supernatant, using p24 ELISA to monitor viral replication. Peptide: As discussed above, peptide D consisting of D-amino acid compared to peptide L composed of L-amino acid has more in serum. Long-term anti-HIV activity. Interestingly, peptide 1 consisting of L-amino acid does not decompose in serum (ie, after 4 hours at 37 ° C), but instead binds tightly (maintains interaction in sds gel) to unknown Serum molecules (data not shown). Such tight binding may result in attenuation of the anti-HIV activity of the L-amino acid peptide in serum over time. Thus, in the subsequent study, the D form of peptide 1 was excluded. The peptide was dissolved in DMSO and subsequently diluted in RpjyQ or dmEM 10 in the absence of serum. Transfer-transfer assay: PGEC was seeded at a cell density of 105 cells/well on the surface of the transfer pores of collagen I and fibrin glue coated with a diameter of 2 mm and a pore size of 3 μm. Joined. Feed the insert every 2 days. The single layer on the filter effectively divides the hole into 15 a top chamber and a bottom side chamber. The integrity of each cell monolayer was determined using an epithelial voltmeter/ohmmeter (Millipore). In order to ensure the integrity of the PGEC barrier, the transepithelial electrical resistance of each cell monolayer was monitored, and the extracellular marker inulin was measured by the cell-by-cell permeability coefficient, such as B〇bardt et al., J Virol 81, 395- As described in 405 (2007), the coefficient of penetration was determined by diffusion 2 〇 assay using 14 C-carboxylated inulin (molecular weight 5 〇〇〇, Sigma). After confirming the integrity of the single layer on the Transwell filter,? 〇£0 exposed to 111¥ (added to the upper room), by borrowing? 24 This 18 eight measures the amount of viral shells to monitor the release of HIV into the basement. In particular, HIV-free cells (10 ng with or without NL4.3g pl60 hypothetical NLd 149 200837075

Env p24) or cell-bound virus (co-cultured with 104 CCR5 + Jaccarat cells 80 μl) was added to the upper chamber for 8 hours at 37 ° C in the presence or absence of 5 μ of peptide 1 . The amount of virus transported by the cell was quantified by the p24 ELISA in the chamber corresponding to the bottom surface. 5 In Vitro Infection and Transmission: Healthy skin was obtained from a full-scale operation, and a dermatome was cut into 3 mm thick slices using a skin slicer within 3 hours after surgery. Divided skin with 1 mg/ml dispase Π (Roche Diagnostics (Pandzburg, Germany) in complete IMDM (Iskov (lSC0ves) modified Dubec medium (IMDM), 10% FCS and 10 μg/ml Gentamicin 10 (gentamycine) was co-cultured for 18 hours at 4° C. The dermal tissue and epidermal tissue were mechanically separated and cut into 1 cm square pieces. The epidermal sheets floated on 200 μl of IMDM in 24-well plates. The epidermis is flanked upwards. Subsequently, the epidermal sheet is infected with HIV NL4.3-BaL-eGFP (100 Ng ρ24) by dropping the virus under the lamella. 1 〇μΜ peptide 1 or an appropriate volume of 15 DMSO control The solution was added in a total volume of 500 μl. After 2 hours, 1.5 ml of complete IMDM was added and the sheets were cultured for 3 days. The epidermal sheets were removed and 200,000 CCR5 + Zhukaert cells were added for another 4 days. The migrated epidermal cells (3rd) and Co-culture samples (grades 5 and 7) were harvested, fixed in 4% tripolyformic acid/PBS, and flow cytometry was used to analyze the expression of GFP. To determine the phenotype of infected migrating cells, the cells were LC labeling Langerin double All infected cells were positive for Langjilin (data not shown). After the addition of peptide 1, the flow cytometry (data not shown) was measured for LC migration, LC maturation and LC survival. DC transmission assay: DCs were seeded at 950 cells/well at 96 150 200837075 . 5 • Well V-shaped bottom plate. Cells with HIV NL4.3-eGFP (X4), NL4.3-BaL-eGFP (R5) Or a single round of NL4.3 △ Env-Egfp (X4) (25 ng p24) with NL4.3 gpl60 hypothetical type for 2 hours at 37 ° C. The cells were washed 3 times with warm medium and added with PHA/IL- 2-activated CD4+ T cells (200,000). Cells were cultured in flat-bottom 96-well plates, harvested 3 days later, fixed in 4% paraformaldehyde/PBS, and GFP expression was determined by FACS. Cytotoxicity: PGEC inoculated in clear bottom 96 Aperture plates. Cells were serially diluted from 55,000 cells to 25 cells in 100 μl of complete DMEM. Add 15 μl of Cell Quantification (CellQimnti)-MTTTM Reagent (Gentaur Belgium), cells at 37 Incubate for 4 hours at ° C. Then, 100 μl of the solubilizing solution was added, and the well plate was shaken at room temperature for 1 hour. OD 570 nm was measured on a Molecular Devices (Molecular Devices) spectrum maximum (SpectraMax) 384 Plath (Plus) reader. A linear relationship was observed between OD 570 nm and cell number. From the blank control group, the detection limit was estimated to be 950 cells. The cytotoxicity of the peptide to PGEC was determined for 15 I, and 55,000 cells were seeded in each of the 80 microliter wells of the clear bottom 96-well plate. The cells were treated twice with 200 μM peptide 1 or 0.01% saponin for 1 week. The cells were continuously exposed to the peptide without washing. 20 Virus speed sedimentation assay: The virus was microcentrifuged at 4 °C for 90 minutes to remove the free shell and resuspended in PBS' to expose the peptide to various conditions and load onto a 20-70% sucrose gradient. After the SW41 T rotor was ultracentrifuged at 20,000 impressions for 24 hours, each fraction was collected and tested for viral protein content. The HIV-1 capsid was assayed by p24 ELISA or by using anti-shell IgG obtained by the AIDS Research and Reference Program by immunoblotting. Hiv-i 151 200837075 gp41 is tested by the anti-____?41 IgG obtained by the AIDS Research and Reference Program by immunoblotting. The RT system is analyzed by the 〇1^ test analysis 20 as described above. Briefly, an entire portion (10 microliters) of each fraction was mixed with an RT reaction mixture (20 microliters). The reaction mixture was incubated at 37 ° C for 2 hours, tapped on a DE81 filter, washed twice with 2xSSC (lx SSC of 0·15 M NaCn, 0.015 M sodium citrate), and once with 95% ethanol. The scintillation count was quantified for 3 h. The density of each sucrose gradient fraction is determined by measuring the refractive index as previously described. Attachment and internalization assay analysis: Attachment and internalization assays are performed as described in 10 Bobardt et al., J Virol 81, 395_405 (2007). For virus attachment, TZM cells (5 〇〇, 000) were exposed to 1 ng of P24 virus for 1 hour, thoroughly stripped to remove unbound virus and dissolved. Under these conditions, no disease is internalized into the cell. For virus internalization, TZM cells (5(9), 〇〇〇) were incubated on 1C for 1 hour at 37C, washed and trypsinized to remove the attached virus and dissolve. The number of attached viruses and internalized viruses in cell lysates was determined by p24 ELISA. Results The results are summarized in the table below. 152 200837075

Table 18A. Broad peptide antiviral effective range Protein name SEQ ID NO peptide sequence IC5〇μΜ IC9〇μιη Host protein apolipoprotein Α-Ι 120 DWLKAFYDKVAEKLKEAF 87.99 145.87 buffalin (buforin) 148 TRSSRAGLQWPVGRVHRLLRK >500 >500 Cathelicidin (LL-37) 149 LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPKrES >500 >500 cynthaurin 150 ILQKAVLDCLKAAGSSLSKAArrAIYNKir >500 >500 dermaseptin 151 GLRSKIWLWVLLMIWQESNKFKKM 68.31 102.11 Haicate 152 FALALKALKKALKKLKKALKKAL >500 >500 K taarcin 153 SMWSGMWRRKLKKLRNALKKKLKGE >500 >500 merittin 154 GIGAVLKVLTTGLPALISWIKRKRQQ 93.56 134.23 PGLa 155 GMASKAGAIAGKIAKVALKAL >500 >500 car xiding (piscidin) 156 FFHHIFRGIVHVGKIIHRLVTG &gt ;500 >500 disease Qin protein P HIVgp41 157 YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF 0.55 0.88 HCVNS5A (peptide 1) 43 SWLRDIWDWICEVLSDFK 1.31 7.97 HIV-1 Vpr 158 EPYNEWTLELLEELKSEAVRH >500 >500 FIV surface glycoprotein 159 EGPTLGNWAREIWArLFKKA >500 >500 Influenza virus hemagglutinin 160 GLFGAIAGFIENGWEGMIDG >500 >500 Influenza virus M2 161 PLWAASnGIIJBLILWIL >500 >500 SFV replicase 162 GSTLYTESRKLLRSWHLPSV >500 >500 Get the table The results of 18(a) are as follows. The sputum cells were exposed to j 5 NL 4.3 in the presence of increasing amounts of amphiphilic peptide derived from the host protein or viral protein. The virus and peptide were washed off after 4 hours. The infection was determined by galactosidase activity 48 hours after infection. 153 200837075 Table 18B Antiviral effective range of peptide 1 HIV-1 name peptide 1 Non-amphiphilic peptide monoclonal IC5〇μΜ IC90 μΜ IC5〇μΜ IC9〇μΜ Single branch system using co-receptors 0.78 4.3 >500 >500 92RW021 A R5 1.2 5.6 >500 >500 92UG029 A X4 0.5 2.3 >500 >500 92TH026 B R5 2.4 7.1 >500 >500 92HT599 B X4 1.8 4.3 >500 &gt ;500 93IN101 C R5 0.7 2.2 >500 >500 98IN017 C X4 1.7 3.9 >500 >500 92UG005 D R5 0.3 1.1 >500 >500 92UG024 D X4 2.4 7.4 >500 >500 92TH006 E R5 1.8 4.6 >500 >500 93TH053 E X4 1.1 3.2 >500 >500 93BR029 F R5 2.1 7.4 >500 >500 93BR020 F X4 0.7 1.8 >500 >500 RU132 G R5 0.9 2.5 >500 > 500 Jvl083 G R5 Drug-resistant HIV-1 1.7 5.2 >500 >500 RT74V 1.2 4.2 >500 >500 RTM184V 1.1 3.1 >500 >500 P Rm461/L63P/V82T/1 84V 2.5 6.3 >500 &gt ;500 PRG48V/L90M 0.7 2.3 >500 >500 T2〇v38a/N42D 1.2 3.5 >500 >500 T2〇N42S Other anti-recording virus 1.6 5.8 >500 >500 SIVmac251 32H 2.7 6.8 >500 >500 SIVsykl.2 1.4 4.3 >500 >500 HIV-2CDC310342 2.6 6.8 >500 >500 HIV-27312A 0.9 2.3 >500 >500 SHIVSF162P3 other virus >500 >500 >500 >500 Adenovirus>500 >500 >500 >500 Vesicular stomatitis virus 154 200837075 For Table 18(b), the target cell line is in increasing number Wild type peptide 1 (SWLRDIWDWICEVLSDFK (SEQ ID NO: 43) or its non-amphibious variant (SWRLDIWDWICESVLDFK, SEQ ID NO: 119), which is exposed to viruses in the absence of antiviral activity (HIV-:l, HIV-2) , SIV, 5 SHIV, Adeno-GFP or VSV-GFP). TZM infection was scored by /3 galactosidase activity (HIV and SIV) or GFP content (VSV-GFP), and 293 infection was scored by GFP content (Adeno-GFP). The results represent three separate experimental results. The results (repeated three times) are expressed as the desired peptide concentration (μΜ) which inhibits the viral infectivity of 50% (IC5〇) or 90% (IC9G). 1〇The results indicate that most of the host-derived peptides including Hector, Sacine, Pistin, PGLa, Latasin-1, Buffalo 2, LL-37 do not affect HIV infection, but are derived from apo The peptides of protein AI, demectin and meridine have mild antiviral activity at high concentrations (Table 18a). Similarly, most of the virus-derived peptides such as Semliki forest virus replicase protein _ 丨, epidemic 15 flu virus M2 protein, influenza virus hemagglutinin, jjjvd

Vpr, and cat immunodeficiency virus glycoproteins do not affect the infection (table coffee). In contrast, amphiphilic peptides derived from HIV gp41 and HCV NS5A (peptide 1), previously shown to inhibit HCV infection, were effective in blocking hIV infection (Table 18a). The amphiphilic peptide derived from HIV gp41 corresponds to the FDA-approved τ2 〇 20 peptide, blocking mv infection via binding to 41 and interfering with fusion (see Kilby et al., Nat. Med. 4, 1302_07 ( 1998)). Conversely, the peptide does not correspond to any HIV gene sequence, thus indicating a novel anti-hiv agent. Peptide 1 blocks all Η1%1 isolated isolates from different branches of 155 200837075 using various co-receptors (used by CCR5 or CXCR4) at low micromolar concentrations or submicromolar concentrations (Table 18b), Drug resistant viruses (Table 18b) and other lenticular viruses (HIV-2, SIV and SHIV) (Table 18b). However, peptide 1 did not block the infectivity of adenovirus or vesicular stomatitis virus (VSV) (Table 18b), suggesting that peptide 1 is selective for neutralizing HIV. Peptide 1 blocks HIV infection in three major in vivo targets of HIV: CD4+ T-lymphocytes, macrophages, and DCs (Fig. 16a, insets 1-3). Clearly, peptide 1 also blocks HIV infection and spread by co-culture of DC-T cells (Fig. 16a, Fig. 4). Similarly, 293 T cells that were transiently treated with peptide treatment and washed with HIV transfected only produce non-infectious particles (Fig. 16b), suggesting that peptide 1 neutralizes the surface of the virus (germinated particles) and And intracellular viruses (assembled particles). In addition, peptide 1 blocked the established infection because the replication of HIV was eliminated even after 3 days of infection (Fig. 16a, Fig. 5). 15 peptide 1 does not eliminate HIV infection by interfering with gpl20 or gp41 because peptide 1 inhibits the infectivity of VSV-G-pseudotype HIV (Fig. 16c). In order to better explain the anti-disease function, 'Shengfu 1 is added to the cells before or after HIV addition (different 16d). ^ When the cells were cultured before the peptide 1 and washed before the addition of the virus, no inhibitory effect was observed (Fig. 16d, left panel). Conversely, the peptide was added to the virus 2 hours before the addition of the disease to the virus (not shown), or the peptide was added after 2 hours, and then the peptide was maintained in culture to prevent mv infection (Fig. 16d, right) Figure), suggesting that peptide 1 acts on the virus rather than on the cell. As discussed above, peptide 1 is virulent to HCV and may become unstable via the membrane layer at 156 200837075. Therefore, the impact of peptide 1 on the HIV structure was analyzed. The untreated virus was precipitated at a density of 1.16 g/cm3, such as the presence of the viral capsid, gp41 and RT (Fig. 17a, left panel). Conversely, all viral components, including membrane-bound gp41, were relocated 5 to the top of the gradient when processed (p. 17a, right), suggesting that peptide 1 destroyed the integrity of the viral particles. Confirming this hypothesis, peptide 1 prevents the virus from attaching to cells and internalizing into the cells (Fig. 17b). It is presumed that the peptide 1 forms a pore in the cell membrane, and the physical link between the small end of the cone-shaped shell core and the virus envelope is unstable, and the link is referred to as a core-membrane linkage (CEL). The core is ίο disassembled in the absence of CEL. The pores induced by the skin 1 will promote the osmotic expansion of the virus caused by the imbalance between the inside and the outside of the virus. However, peptide 1 even destroyed HIV in isotonic buffer, and the buffer mimics the solute concentration of the antiviral program (data not shown). Even the sub-micromolar concentration (0.6μΜ) peptide 1 is enough to make HIV unstable (17th: (Figure). Antiviral effect is not temperature dependent, because the peptide works at 4 ° C, 25 The integrity of HIV is unstable at °C and 37 °C (Fig. 17c). The antiviral effect is rapid because HIV is destroyed within 15 minutes (Fig. 17c) and active under acidic conditions (17c) Figure), suggesting that if the acidic environment of the female vagina is used as a microbicide, the peptide 1 is still active in the female vaginal acid ring. The non-amphipathic variant of peptide 1 does not destroy HIV (17d) Figure), confirming that the amphipathicity of peptide i is not critical for its antiviral activity as indicated by the previous ^^^ (Table 18 b). The membrane binding of full-length HCV NS5A protein is restricted to the subset of intracellular membranes. There is no specific receptor for its anchoring. A recent study reported that trypsin 157 200837075 cytoplasmic membrane, disruption indicates the binding of the extended peptide of the intact NS5A membrane (see Cho et al., J. Virol. 2007). Prompt that the protein medium NS5A binds to its labeling membrane in the endoplasmic reticulum. Whether the peptide 1 interacts with the HIV membrane protein, experiments were conducted to determine whether HIV and pre-trypsin culture were 5 to reduce the sensitivity of HIV to be destroyed by peptide 1. It is important that trypsin treatment does not interfere with peptide 1 HIV is unstable (Fig. 17e), suggesting that no trypsin-sensitive molecules on the virus are required for its virucidal activity. Thus, peptide 1 uses trypsin-resistant protein-containing receptors or non-protein-containing receptors (such as lipids). To combat HIV, VSV and a variety of other viruses that are not sensitive to peptide 1 can reflect any of these alternatives. Because HCV and HIV are budded by lipid vesicles, VSV is not, possibly VSV membranes The protein composition and/or lipid composition is different from that of HIV. The NS5A error-fixing functional site in the HIV membrane is rich in specific protein-containing receptors or lipid receptors, which explains why it is not effective for VSV, most importantly, for host cells. Non-toxic. 15 Effective anti-HIV agents will interfere with three different mechanisms involving HIV transmission: epithelial transfection, DC vector transmission, and mucosal target cell infection. To test the antiviral activity of peptide 1 for HIV transocclusion In effect, an in-vitro assay that mimics the migration of HIV through a genital epithelial cell (PGEC) has been developed. Contrary to the virus lacking gpl60 (Dia 18a), HIV is effectively transferred, suggesting that 20 viral glycoproteins are required for transcellular transport. And suggest that the PGEC layer does not allow non-specific transfer of viral particles. Importantly, peptides can prevent cell-free m V translocation and cell-bound HIV translocation (Fig. 18a). The peptide dose is at least 1 to 1 times higher than the dose required for antiviral activity, and has no cell decomposability for PGEC (different figure). Thus, peptide 1 prevented my 158 200837075 from migrating without disturbing the epithelium intact. Langerhans cells (LC) are DC subsets present in epidermal and epithelial tissues, and LC plays a key role in the spread of HIV from the mucosal site to T cells resident in lymphoid tissues. Therefore, the inventors used the in vitro propagation model 5 to mimic the HIV transmission of LC media. The epidermal sheet is infected with HIV in the presence or absence of peptide 1 . After three trips, the migrated LC analysis directly infects or analyzes its ability to transmit HIV to T cells. The peptide blocks LC infection (Fig. 18c) and HIV transmission to T cells (Fig. 18d and Fig. 18e). Thus, peptide 1 can prevent the spread of HIV by LC in vitro. 10 Monocyte-derived DCs are used to determine the role of the subepithelial DC subset. The spread of HIV from DC to T cells may occur or independently occur in response to DC infection. It is important that peptide 1 blocks both HIV transmission pathways (Fig. 18f). In addition, peptide 1 blocks the spread of replication-defective virus, and it is verified that peptide 1 affects the virus captured by DC (Fig. 18f). Thus, peptide 1 interferes with the spread of HIV in three different layers: PGEC transmigration, DC/LC mediator T cell transmission, and direct DC/LC infection. Thus, peptide 1 is an attractive antiviral agent for the treatment or prevention of HIV. The rapid intracellular and extracellular viricidal mode of mv, the stability of low ph, and the ability to prevent transepithelial migration and prevent cell-to-cell spread, making it particularly attractive as a candidate for microbicide force. By the lipid composition that is lightly recognized in the HIV membrane, it is impossible for the peptide 1 to select an escape mutant, and if the peptide 1 combination is selected for use as a drug-resistant mutant, the spread of the drug-resistant mutant can be prevented. Thus, peptide 1 clearly represents a prototype of a new generation of antiviral agents with HIV treatment or prevention prospects. 159 200837075 Example 17: Additional peptides of the invention Additional peptides of the invention and their antiviral activity are shown in the table below. The ec50 value and the EC9G value were determined as described above. SEQID NO : sequence trait net charge ec5〇(μΜ) ec9〇(μΜ) 43 SWLRDIWDWICEVLSDFK L type, parental-2 0.8 8.0 43 SWLRDIWDWICEVLSDFK type D, parental-2 0.34 1.6 79 SWLRDVWDWVCTILTDFK JFH-1 analogue-1 2.1 17.0 80 SWLRDVWDWICTVLTDFK Con-1 Analog-1 3.9 14.5 123 DWLRIIWDWVCSVVSDFK HCV3a Analog-1 0.55 8.0 124 SWLWEVWDWVLHVLSDFK HCV4a Analog-2 7.0 16.0 125 TWLRAIWDWVCTALTDFK HCV5a Analog 0 7.1 13.9 126 SWLRDVWDWVC3VLSDFK HCV6a Analog-1 3.5 16.9 98 KWLCRIWSWISDVLDDFE Hydrophilic Amino Acid Dispersion-2 0.5 4.0 97 SIWRDWVDLICEFLSDWK Water Repellent Amino Acid Dispersion-2 0.40 3.0 131 SWLKEIWEWICDVLSEFR K, IL&D, E Exchange 2 0.67 3.0 132 SWLKDIWDWICEVLSDFR K, R Exchange-2 0.95 8.8 133 SWLKDIWDWICEVLSDFK R to K -2 1.1 10.5 134 SWLREIWEWICDVLSEFK D, E exchange-2 0.70 4.0 135 SWLREIWEWICEVLSEFK D to E -2 1.0 10.8 136 SWLDRIWRWICKVLSRFE R, D and K, E exchange +2 1.1 2.3 137 SWLDDIWDWICEVLSDFE Only R to D and K to E -6 4.7 16.1 138 SWLRRIWRWICKVLSRFK Only DSR and E to K +6 0.59 1.0 127 SWLRDIW DWISEVLSDFK C to S -2 13.5 16.1 139 SWLRDIWDW1REVLSDFK C to R -1 12.5 16.1 140 SWLRDIWDWffiEVLSDFK C to E -3 13.0 16.5 91 SWLRPIWPWICEVLSDFK First 2D to 2 valine +1 10 nd 141 SGSWLRDIWDWICEVLSDFK Extended N-, 2 aa - 2 4.5 12.0 142 GSWLRDIWDWICEVLSDFK a 丄-2 1.1 7.1 143 SWLRDIWDWICEVLSDFK! 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All of the patents and publications mentioned herein are indicative of the skill of those skilled in the art to which the present invention is disclosed. The full text or the full text is stated here. Applicants reserve the right to any and all materials and information in any patent or publicly available literature cited in this specification for inclusion in this specification. The specific methods and compositions described herein are representative of the preferred embodiments and are intended to be illustrative and not limiting. 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 Those skilled in the art will appreciate that various modifications and changes may be made to the invention disclosed herein without departing from the scope and spirit of the invention. The invention as exemplified herein is suitable for implementation without any of the elements or limitations specifically disclosed herein. The methods and processing procedures described herein may be carried out in a different order of steps, and are not necessarily limited to the order of steps recited herein. As used herein and the appended claims, the singular forms "" Thus, for example, the "one antibody" package includes a plurality of (for example, an antibody solution or a tandem antibody preparation) such an antibody. This patent is in no way interpreted as being limited to the examples or embodiments or methods specifically disclosed herein. This patent must not be construed as limited to any reviewer or patenter's statement of any member or employee of the Bureau unless the statement is specifically and unqualified or reserved in the applicant's written language. 15 Terms and representations are for illustrative purposes only and are not intended to be limiting, and are not intended to exclude the use of such terms and expressions to exclude any equivalent features or portions thereof as shown and described, but A number of changes may be made within the scope of the request for the patent. Thus, it is to be understood that while the invention has been particularly described in terms of a preferred embodiment of the invention, the skilled artisan 20 may resort to modifications and variations of the inventive concepts disclosed herein. It is within the scope of the invention as defined by the appended claims. The invention has been described broadly and comprehensively. The narrower categories and subgroups that fall within the scope of the summary description also form part of the present invention. Included herein is a summary of the invention, which is excluded from the broad category by a proviso or a negative limitation, regardless of whether or not the excluded material is specifically recited herein. Other embodiments are within the scope of the following patent application. In addition, when the 4 inch sign of the month of the month or the aMarkush group of the face is used for explanation, the skilled person knows that the book is also explained by the individual members or subgroups of the Markus_ group. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 shows the position of the peptide relative to the HCV polyprotein genotype la (H77 isolates having SEQ ID NO: 1) and the corresponding anti-HCV activity. Thirteen of the peptides tested in the text test inhibited the infectivity by 9% or more. 1〇 Figure 2 shows the peptide 1 (A) with the amino acid sequence SWLRDIWDWICEVLSDFK (SEQ ID NO: 43) with L-amino acid or D-amino acid added to the cell before exposure to the virus The virus can prevent the initiation of HCV infection; (B) end the ongoing hcv infection; (C) inhibit the HCV infection by growing Huh-7 cells; (D) enter the cell and (E) inhibit intracellular HCV particles 15 Sub-infectivity. As shown in two separate experiments, the EC5 of peptide 1 is approximately 300 nM (F and G). Figure 3 is a histogram showing inhibition of HCV infection in Huh_75J cells by various synthetic peptides, including peptide 1 (SEQ ID NO ·· 43) is the most effective inhibitor of viral infection. 2〇 Figure 4A-G shows the inhibition characteristics and stability of peptide 1 (SEQ ID NO: 43). The L-isomer and D-isomer of peptide 1 are highly inhibitory (A). The peptide 1 acts on the virus (B) and is virucidal (C) to HCV by destroying the HCV virion (D). Both the L-isomer and the D-isomer are effective in membrane permeability (E). The D-form of peptide 1 shows enhanced serum stability (F) and slightly 179 200837075 for the lower IC5 〇 (G). Figures 5A-D are the results showing the effective concentration (EC%) and toxicity (IC%) of the L-form and D-form of peptide 1 (SEQ ID NO.43). Figure 5A shows that the percentage of infected Huh-7.5.1 cells is a function of the concentration of the peptide 1 L-form, and 5 shows that the EC% of the peptide 1 L·form is 0.6 micromolar. Figure 5B shows the percentage of infected Huh-7.5.1 cells as a function of the concentration of the peptide 1 D-form (having D_amino acid instead of L-amino acid), showing that the EC50 of the peptide 1 D-form is 1.0 micromolar concentration. Figure 5C shows that the percentage of viable Huh-7 cells is expressed as the L-form of the peptide 1 (square symbol) and the concentration of the D-form (diamond symbol). The number of L-forms is less than the LC% of the L-form. The LC% of the 46 micromolar concentration and the D-form of the skin 1 is 87 micromolar. Figure 5D shows the percentage of viable Huh-7.5·1 cells as a function of the concentration of the L-form (square symbol) and D-form (diamond symbol) of the peptide 1, showing that the LC50 of the L-form of peptide 1 is 64. The micromolar concentration and the D-form of the peptide 1 are LC5G at a concentration of 105 micromolar. 15 Figures 6A-E show the amphiphilic helical nature of peptide 1 (SEQ ID NO..43). The helical wheel diagram of Figure 1 shows the distribution of the amino acid to obtain a hydrophilic (or polar) face and a water repellent (or non-polar) face (Fig. 6A). The results of the circular two-color diagram show the L-isomer of the peptide 1 and the α-helical structure of the D-isomer (Fig. 6B), the dimethylamino naphthalenesulfonate on the peptide 1 L-isomer and The effect of the α-20 helix structure of the D-isomer (Fig. 6C), and the α-helix of the peptide 1 variant with the C-terminal truncation (Fig. 6D) and the 截 end truncation (Fig. 6) structure. The sequences of these truncated peptides are provided in Table 9. Fig. 7 shows the results of the analysis of the liposome release assay obtained for the plurality of truncated mutants of peptide 1 (SEQ ID NO: 43). The sequence of these truncated peptides is described in Table 9 2008. Figure 8 is a line graph showing that peptide 1 (SEQ ID NO: 43; shown as #7 20 8 in this icon) does not block vesicular stomatitis virus (V S V ) infection. Additional studies indicate that peptide 1 cannot block influenza virus (influenza virus), bovine virus, : 5 Borna disease virus, lymphocytic choriomeningitis virus, or adenovirus infection. ^ Figure 9 is a line graph showing the peptide 2022 (peptide 1) having the sequence: SWLRDIWDWICEVLSDFK (SEQ ID NO: 43) and the peptide 2013 having the sequence SWLRDIWDWICEVL (SEQ ID NO · 92) 10 as indicated by the ELISA, It can substantially inhibit 100% dengue virus infection. The peptide 2017 with the sequence LRDIWDWICEVLSDFK (SEQ ID NO: 107) has a slightly lower activity and inhibits dengue virus infection by about 84%. Figure 10A_D is a line graph showing the percentage of dengue virus infections of various peptides. Figure 10A shows dose-dependent inhibition of dengue virus-infected peptide 2022 (peptide _-1; SEQ ID NO: 43), peptide 2013 (SEQ ID NO: 92), and peptide 2017 (SEQ ID NO: 丨〇 7) The role, such as intracellular detection of dengue virus antigen staining cells by FACS analysis. As shown, at a concentration of 20 μΜ, almost 100% of dengue virus infections were inhibited by peptides 2022 (peptide 1) and 20 peptides, as indicated by FACS. The peptide 2017 has a slightly lower activity at 20 μΜ, inhibiting dengue virus infection by up to about 8%. Figure 10B-D also shows that the leptin variant strain inhibits dengue virus infection and verifies the inhibitory effect of the amphiphilic peptide belonging to the peptide 1 variant, such as intracellular 1^(:^ staining detection. It is shown that as long as the amphiphilic nature of the peptide structure is maintained, the variation of peptide 1 181 200837075 strain and homologue can retain the inhibitory activity. Note that peptide 1 (SEQ ID NO: 43) is called peptide in Figure 10B-D. L-7208. Figure 11 before the virus was added to SHuh-7 cells, when peptide 1 (SEQ ID NO: 43; referred to as 2022 in this figure) and peptide 2012 5 (SWLRDIWDWICEVLSD, SEQ ID NO: 94) and infection The result of the ability of these peptides to inhibit West Nile virus (WNV) infection when the virus is co-cultured. In this figure, the virus is added to the Huh-7 cells in the micro-valency plate, and the WNV system is 102 to 105 PFU in advance. Co-culture with DMSO (first column) or peptide 1 (18 micromolar concentration) (third column) or peptide 2012 (fifth column) 10. The seventh column shows uninfected cells. Afterwards, the well plates were stained with WNV-specific antibodies. As shown, the entire monolayer that has been co-cultured with the DMSO-treated virus is easily detected. Viral antigens were detected. Conversely, there were only 4-5 stained cell focuss in cultures inoculated with 105 PFU of peptide-treated virus, whereas there was no evidence of infection in cultures vaccinated with lower doses of virus. 15 Figure 12 The diagram shows that both the L-form and the D-form of peptide 1 are L-2022 [SWLRDIWDWICEVLSDFK] (SEQ ID NO: 43) and D-2022 (SEQ ID NO: 43 with D-amino acid) at 1 • A concentration of 25 micromolar to 5.0 micromolar inhibits HIV-1 infection. Also shown is peptide 2018 (DIWDWICEVLSDFK) (SEQ ID NO: 108) which is a 20-terminal truncated 14-mer of peptide 1 The version has similar antiviral activity; and the peptide 2504 (SWLRDIWDWICEV) (SEQ ID NO: 103) is a C-terminal truncated 13-mer analog of peptide 1 with slightly lower activity. Conversely, peptide 2015 ( SWLRDIWDWI) (SEQ ID NO: 105) is a C-terminal truncated 10-mer version of the peptide 1 that is not active; peptide 6938 182 200837075: 5 • (LYGNEGCGWAGWLLSPRG) (SEQ ID NO: 6) is derived from the HCV core The 18-mer of the protein is also inactive. Figure 13A-B is a diagram showing the live L-7208 HS peptide (SIWRDWVDLICEFLSDWK, SEQ ID NO: 97) Almost equal to the height of the active L-7208 SEQ ID NO: 43 peptide (also known as L-2022 peptides or "peptide 1"). L-7208 HS peptide has the same amino acid composition as L-7208 peptide, but the water-repellent amino acid of L-7208 HS peptide (SEQ ID NO: 97) has been broken up, thereby changing the amine The base acid sequence, but maintains its amphiphilic nature. Thus, L-7208 HS peptide (SEQ ID NO..97), 10 L-7208 peptide (SEQ ID NO..43) and D-7208 peptide (SEQ ID NO: 43 contains D-amino acid) Non-L-amino acids) generally inhibit 100% HIV infection at concentrations of about 20 micromolar. Another peptide (referred to as 3229, SWRLDIWDWICESVLDFK, SEQ ID NO: 119) whose amino acid is exchanged to reduce its amphiphilic properties, but which has a peptide with L-7208 (SEQ ID 15 • A NO : 43) The same amino acid composition, very little, if any, is active. These data indicate that the amphiphilic nature of the peptide is important for the activity, but the exact sequence of the peptide is not particularly limited. The results for HIVR9BaL from 293 T cells are shown in Figure 13A. The results obtained from CEM T cells for HIV R9BaL are shown in Figure 13B. 20 Figure 14A-C shows the peptide in the extracellular metastable ΗΙν_ι virion. For example, Figure 14 shows that after treatment of HIV-1 preparation with peptide 1 (labeled as l_7208 (SEQ ID NO: 43)), the infectious virion releases a large amount of free HIV-1 shell' indicating that the virion is The peptide is dissolved. Conversely, after treatment with HIV-1 virions in DMSO or with control peptide 6938 183 200837075 (LYGNEGCGWAGWLLSPRG) (SEQ ID NO: 6), the HIV-1 shell was not substantially released. Consistent with these observations, the number of shells associated with the virus treated with L-7208 (SEQ ID NO: 43) (Fig. 14B) was reduced, but with the peptide 6938 (LYGNEGCGWAGWLLSPRG, 5 SEQ ID NO: 6) The sample processed is no. Finally, Figure 14C shows the HIV-1 shell internalized into the cell after treatment with DMSO and 5 or 10 micromolar concentration peptide 6938 (SEQ ID NO: 6) or L-7208 (SEQ ID NO: 43). The percentage. When cells were treated at 5 or 10 micromolar concentrations of 6938 (SEQ ID NO: 6), approximately 1% of the control number of HIV-1 capsids were internalized; 10 by the use of peptide L-7208 (SEQ ID) NO: 43) treatment, the internalization of HIV-1 shell was inhibited by 10 times. Figure 15 is a histogram showing that the peptide having an amphiphilic structure similar to the amphipathic structure of peptide L-7208 (SEQ ID NO: 43) can also strongly inhibit HIV infection. Thus, an amphiphilic peptide having potent anti-HIV activity includes a peptide 1522 (SEQ ID NO: 127), a peptide 3226 (SEQ ID NO: 128), a peptide 3228 (SEQ ID NO: 130), a peptide L-7208 2D to 2 Pro (SEQ ID NO: 91) and L-7208 HS carry a dissociated hydrophilic amino acid (KWLCRIWSWISDVLDDFE, SEQ ID NO: 98). Figure 16A-D: peptide 1 (SEQ ID NO: 43; amphipathic "viral 2 sputum") not only neutralizes HIV without cells, but also neutralizes HIV and internalization. HIV. (A) CD4+ T-lymphocytes, macrophages or DCs (0.1 x 16 cells) were exposed to NL4.3 BaL (l-nike p24) 10 曰, washed three times with the culture medium, and cultured in a flat-bottom 96-well plate. Wild-type peptide 1 or its non-amphibious variant (SEQ ID NO: 119; 5 μΜ) was added to CD4+ 184 200837075 T-cell, python and DC (Figs. 1 to 3) together with the virus, and was added before T cell culture. The DC was pulsed by HIV (Fig. 4) or added to the tau cells 3 times after infection (Fig. 5). Supernatants were collected at different days and the virus was monitored by p24 ELISA. The error bars represent the standard deviation of the replicates. These experiments represent three independent experiments using three or five different donors. (B) 293 T cells infected with NL4.3 or NL4.3 BaL for 24 hours were transfected with a peptide (5 μM) at 37 ° C or without using a peptide for 1 hour, and the peptide was removed by washing. The infectivity of the virus released at 293 ΤΖΜ on the sputum cell score 24 hours after the peptide treatment. The infection was determined by the galactosidase activity 48 hours after infection. The error bars represent the standard deviation of the repeated test 10. These experiments represent two independent experiments. (C) TZM cells were exposed to VSVG pseudotype pNL4.3-AEnv virus (1 Nike p24) from gpl60 NL4.3 (X4), gpl60 BaL (R5) or with or without peptide (5 μM). The infection was determined by the 5-galactosidase activity 48 hours after infection. The error bars represent the standard deviation of the replicates. Data are expressed as a percentage of infection. (D) TZM cells 15 were pretreated with peptide (5 μM) for 1 hour, 2 hours, 4 hours, and 8 hours, thoroughly washed to remove the peptide, and then exposed to NL4.3 (1 Ng ρ24); or cell first Exposure to virus and addition of peptides after 1, 2, 4 and 8 hours. The infection was determined by cold-galactosidase activity 48 hours after infection. The error bars represent the standard deviation of the repeated experiments. 20 Figure 17A-E: peptide 1 (SEQ ID NO: 43; amphipathic "viral agent") destroys the membrane core protein of HIV and the core protein of the shell intact. (A) Purified NL4.3 virus (20 ng p24 at ?63) with or without peptide 1 (5 μM) was incubated at 37 ° C for 30 minutes and loaded on a 20-70% curtain sugar gradient. The collected gradients were analyzed for the shell (P24 ELISA and immune dots), 185 200837075 RT (by exoRT assay) and gp41 content (by immunoblots). The starting points and degrees (g/cm3) of each of the selected points shown in the lower two figures are measured by measuring the refractive index. (B) For virus attachment 'TZM cells (500,000) to 4. The 〇 was exposed to 1 ng of P24 NL4.3 for 1 hour and thoroughly washed to remove unbound virus and dissolve. After internalization with the virus, the cells were exposed to the virus for 2 hours at 37 < t, washed, trypsinized to remove the attached virus and dissolved. The amount of virus attached to the internalization virus was determined by cell lysate by P24 ELISA. The error bars indicate the standard deviation of the repeated tests. The data is expressed as a percentage of attachment or a percentage of internalization. These experiments represent two independent experiments. (C) Same as (A), but the virus was treated with a decreasing concentration of peptide 1 at 37 ° C for 30 minutes (top left); (: treated with 5 μΜ peptide 1 for 15, 30 and 60 minutes (bottom left); at 4. 〇, 25. (: or 37. (: 30 minutes of treatment (top right); and ρΗ 8, 7, 6 and 5 treatment for 30 minutes (bottom right panel). Gradient selection of the HIV shell by p24 ELISA. (D) Same as (A), but the virus is treated with peptide 1 or with its non-amphibious variant (SEQ ID NO) : 119) 15 treatment. (E) Same as (A), but the virus was first trypsinized at 37 ° C for 15 minutes, neutralized with trypsin in 10% FCS, and microcentrifuged at 4 ° C for 90 minutes to resuspend. Immediately loaded onto the sucrose gradient for assessing viral integrity by p24 ELISA. Figure 18A-F: peptide 1 (SEQ ID NO: 43; amphipathic "viricidal 20 agent") inhibits HIV reproductive epithelial migration and LC/DC forwarding of HIV. (A) Cell-free HIV or cell-associated HIV was added to the top surface of primary reproductive epithelial cells (PGEC) at 37 °C for 8 hours. The amount of virus transported through the cell was borrowed from the bottom surface of PGEC. Corresponding to the p24 ELISA quantification of the chamber. In order to determine the effect of peptide 1 (SEQ ID NO: 43) on HIV transfer, just add the virus 186 200837075 to PGEC The effect of peptide 1, peptide 1 was compared to its non-amphibious variant (SEQ ID NO: 119). The results were expressed as the percentage of p24 originally inoculated. The error bars represent the standard deviation of the replicates. Representative of 4 independent experiments of PGEC of each donor. (B) PGEC was treated twice with 5 200 μΜ peptide 1 (SEQ ID NO: 43) or 0.01% saponin for 1 week. No washing was performed to maintain the cells. Continuous exposure to the peptide. After overnight incubation, cell quantitation (CellQuanti)-MTTTM reagent was added and cell viability was quantified by OD 570 nm reading. As for the 18 CF map, the epidermal sheet was treated with HIVNL4.3-BaL-eGFP ( 100 ng p24) infection, co-cultured directly with ΙΟμΜ 10 peptide 1 (SEQ ID NO: 43) or DMSO control group. After 3 曰, the epidermal sheet was removed, and 200,000 CCR5 + Jurkat cells were added and further sputum was added. The migrated DC/LC epidermal cells (Day 3) and co-cultured samples (Days 5 and 7) were analyzed by FACS for green fluorescent protein (GFP) expression. (C) The migrated DC/LC HIV infection line was Represented as a percentage of total cells. Error bars indicate the standard of 15 repeated experiments Poor. (D) Co-culture Further analysis of infection by FACS on day 7. The percentage of infected cells was shown. (E) The donor variation rate shown on day 5. The error bars represent the standard deviation of the replicates. (F) DC (50,000) exposed to HIV-1 NL4.3-eGFP (X4), NL4.3-BaL-eGFP (R5) at 37 °C or exposed to NL4.3 gpl60 env (25 Nike p24) Pseudotypes of NL4.3A 20 Env-eGFP together with 1 〇μΜ peptide 1 or DMSO control group for 2 hours. The cells were washed 3 times, activated CD4+ T cells were added for 3 days, and GFP expression was measured by FACS. The error bars represent the standard deviation of the replicates. These results represent 3 independent experiments. [Main component symbol description] (none) 187

Claims (1)

200837075 X. Patent application scope: 1. A separated peptide of 14 to 50 D-amino acids or L-amino acids, comprising the amino acid sequence of formula I: Xaa!-Xaa2'Xaa3_ WL- Xaa6-Xaa7-Xaa8_W-Xaai 〇-W-Xaai 2~Xaa! 3 -Xaai4-Xaai5-Xaai6-Xaai7-Xa3«i8_Xaai9_Xaa2〇-Xaa2i-Xaa22 (SEQIDNO: 163)? Among them 331 is serine (S) Or is absent; Xaa is glycine (G) or absent; Xaas is serine (S), aspartic acid (D) or threonine (T); W is tryptophan; Leucine; Xaa0 is arginine (R), aspartic acid (D), lysine (κ), tryptophan, or glutamic acid (E); Xaa? is aspartic acid (D), fine Aminic acid (R), glutamic acid (E), alanine (A), isoleucine (I), or lysine (K); Xaas is isoleucine (1) or proline (v); Xaa1G is aspartic acid (D), arginine (R), glutamic acid (prone or lysine; Xaan is isoleucine (1) or valine (v); Xaan is cysteine (C) , glutamic acid, leucine (L), serine (s) or arginine (R); XaaM is amylin (6), lysine (κ), aspartic acid (9), threonine (τ ), histidine (H), serine (s) or arginine (R); Xaal s is amylin (v), alanine (A) or serine (8); 200837075 and & 316 is leucine (L) or valine (V); Xaan is serine (S), S Amine acid (T), leucine (L) or absent; Xaals is aspartic acid (D), arginine (R), glutamic acid (E), lysine (K) or non-existent Xaai9 is phenylalanine or is absent; Xaa2 is lysine (K), glutamic acid (E), arginine (R), aspartic acid (D) or is absent, Xaa2i is stepped on Amine acid (T) is either absent; and Xaa22 is tryptophan (W) or absent; and wherein the amino acid sequence of formula I is: SWLRDIWDWICEVLSDFK (SEQ ID NO: 43); SWLRDIWDWICEVLSDF (SEQ ID NO) : 95), SWLRDIWDWICEVLSD (SEQ ID NO: 94), SWLRDIWDWICEVLS (SEQ ID NO: 93), or SWLRDIWDWICEVL (SEQ ID NO: 92). 2. The isolated peptide of claim 1 of the patent scope, An amino acid sequence comprising a group selected from the group consisting of: SWRLDIWDWICESVLDFK (SEQ ID NO: 119), DWLRII WDWVCSVVSDFK (SEQ ID NO: 123), SWLWEVWDWVLHVLSDFK (SEQ ID NO: 124) 5 TWLRAIWDWVCTALTDFK (SEQ ID NO) : 125), SWLRDVWDWVCTVLSDFK (SEQ I D NO: 126), SWLRDIWDWISEVLSDFK (SEQ ID NO: 127)? SWLDRIWRWICKVLSRFE (SEQ ID NO: 128), SWLDDIWDWICEVLSDFE (SEQ ID NO: 129), SWLRRIWRWICKVLSRFK (SEQ ID NO: 130), SWLKEIWEWICDVLSEFR (SEQ ID NO: 131) SWLKDIWDWICEVLSDFR (SEQ ID NO: 132), 200837075 SWLKDIWDWICEVLSDFK (SEQ ID NO: 133), SWLREIWEWICDVLSEFK (SEQ ID NO: 134), SWLREIWEWICEVLSEFK (SEQ ID NO: 135), SWLDRIWRWICKVLSRFE (SEQ ID NO: 136)? SWLDDIWDWICEVLSDFE (SEQ ID NO: 137)? SWLRRIWRWICKVLSRFK (SEQ ID NO: 138), SWLRDIWDWIREVLSDFK (SEQ ID NO: 139), SWLRDIWDWIEEVLSDFK (SEQ ID NO: 140), SGSWLRDIWDWICEVLSDFK (SEQ ID NO: 141), GSWLRDIWDWICEVLSDFK (SEQ ID NO: 142) , SWLRDIWDWICEVLSDFKT (SEQ ID NO: 143), SWLRDIWDWICEVLSDFKTW (SEQ ID NO: 144), SWRLDIWDWICESVLDF (SEQ ID NO: 189), SWRLDIWDWICESVLD (SEQ ID NO: 190), SWRLDIWDWICESVL (SEQ ID NO: 191), SWRLDIWDWICESV (SEQ ID NO: 192), DWLRII WDWVCSVVSDF (SEQ ID NO: 193), DWLRII WDWVCSVVSD (SEQ ID NO: 194), DWLRII WDWVCSVVS (SEQ ID NO: 195), DWLRII WDWVCSVV (SEQ ID NO: 196), SWLWEVWDWVLH VLSDF (SEQ ID NO: 197), SWLWEVWDWVLHVLSD (SEQ ID NO: 198), SWLWEVWDWVLHVLS (SEQ ID NO: 199), SWLWEVWDWVLHVL (SEQ ID NO: 200)? TWLRAIWDWVCTALTDF (SEQ ID NO: 201), TWLRAIWDWVCTALTD (SEQ ID NO) : 202), TWLRAIWDWVCTALT (SEQ ID NO: 203), TWLRAIWDWVCTAL (SEQ ID NO: 204), SWLRDVWDWVCTVLSDF (SEQ ID NO: 205), SWLRDVWDWVCTVLSD (SEQ ID NO: 206), SWLRDVWDWVCTVLS (SEQ ID NO: 207), SWLRDVWDWVCTVL (SEQ ID NO: 208), SWLRDIWDWISEVLSDF (SEQ ID NO: 209), SWLRDIWDWISEVLSD (SEQ ID NO: 210), SWLRDIWDWISEVLS (SEQ ID NO: 211), 3 200837075 SWLRDIWDWISEVL (SEQ ID NO: 212), SWLDRIWRWICKVLSRF (SEQ ID NO: 213), SWLDRIWRWICKVLSR (SEQ ID NO: 214)? SWLDRIWRWICKVLS (SEQ ID NO: 215), SWLDRIWRWICKVL (SEQ ID NO: 216), SWLDDIWDWICEVLSDF (SEQ ID NO: 217)? SWLDDIWDWICEVLSD (SEQ ID NO: 218), SWLDDIWDWICEVLS (SEQ ID NO: 219), SWLDDIWDWICEVL (SEQ ID NO: 220), SWLRRIWRWICKVLSRF (SEQ ID NO: 221), SWLRRIWRWICKVLSR (SEQ ID NO: 222), SWLRRIWRWICKVLS (SEQ ID NO: 223), SWLRRIWRWICKVL (SEQ ID NO) : 224)? SWLKEIWEWICDVLSEF (SEQ ID NO: 225), SWLKEIWEWICDVLSE (SEQ ID NO: 226), SWLKEIWEWICDVLS (SEQ ID NO: 227), SWLKEIWEWICDVL (SEQ ID NO: 228), SWLKDIWDWICEVLSDF (SEQ ID NO: 229), SWLKDIWDWICEVLSD (SEQ ID NO: 230), SWLKDIWDWICEVLS (SEQ ID NO: 231), SWLKDIWDWICEVL (SEQ ID NO: 232), SWLKDIWDWICEVLSDF (SEQ ID NO: 233), SWLKDIWDWICEVLSD (SEQ ID NO: 234), SWLKDIWDWICEVLS (SEQ ID NO: 235), SWLKDIWDWICEVL (SEQ ID NO: 236), SWLREIWEWICDVLSEF (SEQ ID NO: 237), SWLREIWEWICDVLSE (SEQ ID NO: 238), SWLREIWEWICDVLS (SEQ ID NO: 239)? SWLREIWEW1CDVL (SEQ ID NO: 240), SWLREIWEWICEVLSEF ( SEQ ID NO: 591), SWLREIWEWICEVLSE (SEQ ID NO: 592), SWLREIWEWICEVLS (SEQ ID NO: 593), SWLREIWEWICEVL (SEQ ID NO: 594), SWLDRIWRWICKVLSRF (SEQ ID NO: 241), SWLDRIWRWICKVLSR (SEQ ID NO: 242 ), 4 SWLDRIWRWICKVLS (SEQ ID NO: 243), SWLDRIWRWICKVL (SEQ ID NO: 244), SWLDDIWDWICEVLSDF (SEQ ID NO: 245), SWLDDIWDWICEVLSD (SEQ ID NO: 246), SWLDDIWDWICEVLS (SEQ ID NO: 247) 5 SWLDDIWDWICEVL ( SEQ ID NO: 248), SWLRRIWRWICKVLSRF (SEQ ID NO: 249), SWLRRIWRWICKVLSR (SEQ ID NO: 250), SWLRRIWRWICKVLS (SEQ ID NO: 251), SWLRRIWRWICKVL (SEQ ID NO: 252), SWLRDIWDWIREVLSDF (SEQ ID NO: 253), SWLRDIWDWIREVLSD (SEQ ID NO: 254)? SWLRDIWDWIREVLS (SEQ ID NO: 255), SWLRDIWDWIREVL (SEQ ID NO: 256)? SWLRDIWDWIEEVLSDF (SEQ ID NO: 257), SWLRDIWDWIEEVLSD (SEQ ID NO: 258), SWLRDIWDWIEEVLS (SEQ ID NO) : 259), SWLRDIWDWIEEVL (SEQ ID NO: 260), SGSWLRDIWDWICEVLSDF (SEQ ID NO: 261), SGSWLRDIWDWICEVLSD (SEQ ID NO: 262), SGSWLRDIWDWICEVLS (SEQ ID NO: 263), SGSWLRDIWDWICEVL (SEQ ID NO: 264), GSWLRDIWDWICEVLSDF (SEQ ID NO: 265), GSWLRDIWDWICEVLSD (SEQ ID NO: 266), GSWLRDIWDWICEVLS (SEQ ID NO: 267), GSWLRDIWDWICEVL (SEQ ID NO: 268), SWLRDIWDWICEVLSDFK (SEQ ID NO: 269), SWLRDIWDWICEVLSDF (SEQ ID NO: 270), SWLRDIWDWICEVLSD (SEQ ID NO: 271), SWLRDIWDWICEVLS (SEQ ID NO: 272), SWLRDIWDWICEVLSDFKT (SEQ ID NO: 273)? SWLRDIWDWICEVLSDFK (SEQ ID NO: 274), SWLRDIWDWICEVLSDF (SEQ ID NO: 275), and SW LRDIWDWICEVLSD (SEQ ID NO: 276). 200837075 3. The isolated peptide according to claim 1 of the patent application, which is an amine selected from the group consisting of SEQ ID NO: 119, 123-144, 189-276 and 591 face 594. The composition of the base acid sequence. 4. The isolated peptide of claim 1 which comprises an amino acid sequence of formula η: Xaa1-Xaa2-SWL-Xaa6-Xaa7-IW-Xaai〇-WIC- Xaai4-VL-Xaai7 -Xaai8-Xaai9-Xaa2〇-Xaa2i-Xaa22 (SEQ ID NO: 164), wherein: Xaa!, Xaa2, Xaa21AXaa22 are absent; S is serine; w is tryptophan; l is leucine; Xaa0 Is arginine (R), aspartic acid (D), glutamic acid or lysine Xaa7 is arginine (R), aspartic acid (D) or lysine (κ); I is isoleamine Acid; W is tryptophan; XaaiG is aspartic acid (D), arginine (R), or lysine (K); C is hemi-deaminic acid; V is valine (V); Amino acid (K), arginine (R), face acid (6) or aspartic acid (9) Xaan is serine acid (S) or is absent; Xaai8 is aspartic acid (D), fine (8), amine _ or for non-existence; Xaa19 is stupid propylamine brewed. Xaa2 is lysine (κ), face amine or is absent. The peptide comprising one of the following amino acid sequences selected from the group consisting of: 200837075, as set forth in claim 4: SWLEKIWKWICRVLSKFD (SEQ ID NO: 165); SWLRKIWKWICEVLSDFK (SEQ ID NO: 166); SWLRDIWDWICKVLSKFK (SEQ ID NO: 167); SWLRRIWRWICEVLSDFK (SEQ ID NO: 168); SWLRDIWDWICRVLSRFK (SEQ ID NO: 169); SWLRRIWDWICRVLSDFK (SEQ ID NO) SWLRKIWDWICKVLSDFK (SEQ ID NO: 171); SWLRRIWDWICEVLSRFK (SEQ ID NO: 172); SWLRKIWDWICEVLSKFK (SEQ ID NO: 173); SWLRDIWRWICRVLSDFK (SEQ ID NO: 174); SWLRDIWKWICKVLSDFK (SEQ ID NO: 175); SWLDRIWDWICRVLSRFK (SEQ ID NO: 176); SWLRDIWDWICKVLSKFK (SEQ ID NO: 177); SWLEKIWKWICRVLSKF (SEQ ID NO: 393); SWLEKIWKWICRVLSK (SEQ ID NO: 394); SWLEKIWKWICRVLS (SEQ ID NO: 395); SWLEKIWKWICRVL (SEQ ID NO: 396); SWLRKIWKWICEVLSDF (SEQ ID NO: 397); SWLRKIWKWICEVLSD (SEQ ID NO: 398); SWLRKIWKWICEVLS (SEQ ID NO: 399); SWLRKIWKWICEVL (SEQ ID NO: 400); SWLRDIWDWICKVLSKF (SEQ ID NO: 401); SWLRDIWDWICKVLSK ( SEQ ID NO: 402); SWLRDIWDWICKVLS (SEQ ID NO: 403); SWLRDIWDWICKVL (SEQ ID NO: 404); SWLRRIWRWICEVLSDF (SEQ ID NO: 405); SWLRRIWRWICEVLSD (SEQ ID NO: 406); SWLRRIWRWICEVLS (SEQ ID NO: 407 ); SWLRRIWRWICEVL (SEQ ID NO: 408); SWLRDIWDWICRVLSRF (SEQ ID NO: 409); SWLRDIWDWICRVLSR (SEQ ID NO: 410); SWLRDIWDWICRVLS (SEQ ID NO: 411); SWLRDIWDWICRVL (SEQ ID NO: 412); 7 200837075 SWLRRIWDWICRVLSDF (SEQ) ID NO: 413); SWLRRIWDWICRVLSD (SEQ ID NO: 414); SWLRRIWDWICRVLS (SEQ ID NO: 415); SWLRRIWDWICRVL (SEQ ID NO: 416); SWLRKIWDWICKVLSDF (SEQ ID NO: 417); SWLRKIWDWICKVLSD (SEQ ID NO: 418) SWLRKIWDWICKVLS (SEQ ID NO: 419); SWLRKIWDWICKVL (SEQ ID NO: 420); SWLRRIWDWICEVLSRF (SEQ ID NO: 421); SWLRRIWDWICEVLSR (SEQ ID NO: 422); SWLRRIWDWICEVLS (SEQ ID NO: 423); SWLRRIWDWICEVL (SEQ ID NO: 424); SWLRKIWDWICEVLSKF (SEQ ID NO: 425); SWLRKIWDWICEVLSK (SEQ ID NO: 426); SWLRKIWDWICEVLS (SEQ ID NO: 427); SWLRK1WDWICEVL (SEQ ID NO: 428); SWLRDIWRWICRVLSDF (SEQ ID NO: 429); SWLRDIWRWICRVLSD (SEQ ID NO: 430); SWLRDIWRWICRVLS (SEQ ID NO: 431); SWLRDIWRWICRVL (SEQ ID NO: 432); SWLRDIWKWICKVLSDF (SEQ ID NO: 433); SWLRDIWKWICKVLSD (SEQ ID NO: 434); SWLRDIWKWICKVLS (SEQ ID NO) : 435); SWLRDIWKWICKVL (SEQ ID NO: 436); SWLDRIWDWICRVLSRF (SEQ ID NO: 437); SWLDRIWDWICRVLSR (SEQ ID NO: 438); SWLDRIWDWICRVLS (SEQ ID NO: 439); SWLDRIWDWICRVL (SEQ ID NO: 440); SWLRDIWDWICKVLSKF (SEQ ID NO: 441) SWLRDIWDWICKVLSK (SEQ ID NO: 442); SWLRDIWDWICKVLS (SEQ ID NO: 443); and SWLRDIWDWICKVL (SEQ ID NO: 444). 6. The isolated peptide according to item 4 of the patent application, which comprises an amino acid sequence 8 200837075 selected from the group consisting of SEQ ID NO: 165-177 and 393-444. . 7. The isolated peptide of claim 4, which comprises an amino acid sequence of formula m: XaarXaa2-SW-LRXaa7-IW-Xaa10.WI-C- Xaai4-VL-Xaa17-Xaa18 .Xaai9-Xaa2〇-Xaa2i-Xaa22 (SEQ id NO: 178)? Where: Xaa, Xaa2, Xaa2i and Xaa 2 are absent; S is serine; W is tryptophan; L is leucine R is arginine; I is isoleucine; C is cysteine; V is valine; Xaa? is aspartic acid (D), arginine (R) or lysine (κ) Xaa1G is arginine (R) or lysine (κ); XaaH is lysine (K), glutamic acid (E) or aspartic acid (D); \&&17 is a serine (S) is either absent; Xaals is arginine (R), lysine (κ) or absent; Xaa19 is phenylalanine (F) or is absent; and Xaa2 is lysine ( K) or does not exist. 8. The isolated peptide according to item 7 of the patent application, comprising an amino acid sequence selected from the group consisting of: SWLRDIWRWICKVLSRFK (SEQ ID NO: 179); SWLRDIWKWICKVLSKFK (SEQ ID NO) SWLRKIWKWICEVLSKFK (SEQ ID NO: 181); SWLRRIWRWICEVLSRFK (SEQ ID NO: 182); SWLRRIWRWICDVLSRFK (SEQ ID NO: 183); SWLRDIWRWICKVLSRF (SEQ ID NO: 445); SWLRDIWRWICKVLSR (SEQ ID NO: 446); 200837075 SWLRDIWRWICKVLS (SEQ ID NO: 447); SWLRDIWRWICKVL (SEQ ID NO: 448); SWLRDIWKWICKVLSKF (SEQ ID NO: 449); SWLRDIWKWICKVLSK (SEQ ID NO: 450); SWLRDIWKWICKVLS (SEQ ID NO: 451); SWLRDIWKWICKVL (SEQ ID NO: 452); SWLRKIWKWICEVLSKF (SEQ ID NO: 453); SWLRKIWKWICEVLSK (SEQ ID NO: 454); SWLRKIWKWICEVLS (SEQ ID NO: 455); SWLRKIWKWICEVL (SEQ ID NO: 456); SWLRRIWRWICEVLSRF (SEQ ID NO: 457); SWLRRIWRWICEVLSR (SEQ ID NO: 458); SWLRRIWRWICEVLS (SEQ ID NO: 459); SWLRRIWRWICEVL (SEQ ID NO: 460); SWLRRIWRWICDVLSRF (SEQ ID NO: 461); SWLRRIWRWICDVLSR (SEQ ID NO: 462); SWLRRIWRWICDVLS (SEQ ID NO) : 463); and SWLRRIWRWICDV L (SEQ ID NO: 464). 9. The isolated peptide of claim 7 which consists of one amino acid sequence selected from the group consisting of SEQ ID NOs: 179-183 and 445-464. 10. The isolated peptide of claim 4, which comprises an amino acid sequence of formula IV: Xaai-Xaa2_S-WL-Xaa6-Xaa7_I-W-Xaai〇_WIC-Xaai4_V-L-Xaai7_Xaai 8-Xaai9_Xaa2〇-Xaa2i (SEQ ID NO: 184), wherein: Xaai, Xaa2, Xaa2i and Xaa22 are absent; S is serine; W is tryptophan; L is leucine; I is isoleamine Acid; 10 200837075 C is cysteine; hydrazine is _-amino acid; Xaa6, Xaa7, Xaa1 () and Xaa14 are arginine (8) or lysine (κ); Xaan is serine (S) or not Exist; Xaals is arginine (8), lysine (κ) or is absent; Xaa 9 is phenylalanine (smart or absent; and Xaa2 is arginine (R), lysine ( K) or is absent. 11. The isolated peptide according to the first aspect of the patent application, comprising an amino acid sequence selected from the group consisting of: SWLRKIWKWICKVLSKFK (SEQ ID NO: 185); SWLRRIWRWICRVLSRFK (SEQ ID NO: 186); SWLRRIWRWICRVLSRFR (SEQ ID NO: 187); SWLKKIWKWICKVLSKFK (SEQ ID NO: 188); SWLRKIWKWICKVLSKF (SEQ ID NO: 465); SWLRKIWKWICKVLSK (SEQ ID NO) : 466); SWLRKIWKW1CKVLS (SEQ ID NO: 467); SWLRKIWKWICKVL (SEQ ID NO: 468); SWLRRIWRWICRVLSRF (SEQ ID NO: 469); SWLRRIWRWICRVLSR (SEQ ID NO: 470); SWLRRIWRWICRVLS (SEQ ID NO: 471); SWLRRIWRWICRVL (SEQ ID NO: 472); SWLRRIWRWICRVLSRF (SEQ ID NO: 473); SWLRRIWRWICRVLSR (SEQ ID NO: 474); SWLRRIWRWICRVLS (SEQ ID NO: 475); SWLRRIWRWICRVL (SEQ ID NO: 476); SWLKKIWKWICKVLSKF (SEQ ID NO: 477); SWLKKIWKWICKVLSK (SEQ ID NO: 478); SWLKKIWKWICKVLS (SEQ ID NO: 479); and SWLKKIWKWICKVL (SEQ ID NO: 480) ° 12. The isolated peptide of claim ii, It consists of a sequence of amino acid 11 200837075 selected from the group consisting of SEQ ID NOs: 185-188 and 465-480. 13. The virus-killing activity is as described in the patent application. 14. The isolated peptide of claim 1 of the patent application, which is 18 to 40 K amino acids or L-amino acids. 15_ The isolated peptide of claim 1 of the patent scope, which is 18 to 30 amino acids or L-amino acid. 16. The isolated peptide according to item 1 of the patent application, which is 18 to 22 D-amino acids or L-amino acids. 17. A pharmaceutical composition comprising the isolated peptide of claim 1 of the scope of the patent application. 18. The pharmaceutical composition of claim 17, which is a microbicide. 19. A pharmaceutical composition according to claim 17 of the patent application, which is a vaginal cream. 20. The pharmaceutical composition of claim 17, further comprising an anti-viral agent. 21. The pharmaceutical composition of claim 2, wherein the antiviral agent is a protease inhibitor, a polymerase inhibitor, a ligase inhibitor, an invasion inhibitor, an assembly/secretion inhibitor, a translation inhibitor, or Immunostimulant. 22. The pharmaceutical composition of claim 20, wherein the antiviral agent is alpha interferon, pegylated interferon, ribavirin, amantadine, limantadine ( Rimantadine), pleconaril, acyclovir, zidovudine, lamivudine, Indenavir (Merck) ), telaprivir (Fortes 12 200837075 (Vertex)), Tenofivir (Gilead), r1626 (Roche), GS-9137 (Giri) Company), Fuzeon (Roche, Trimeris), celg0sivir (Migenix), VGX-410c (VGX Pharmaceuticals), 〇IMO-2125 (Indira pharmaceuticals) or a combination thereof. 23. A manufactured article comprising a container for collecting a single liquid and one of the peptides of the first item of the patent application. 24. The article of claim 23, wherein the container is a collection bag, a tube, a capillary or a syringe. 25. The article of claim 23, wherein the container is evacuated. 26. If the object of claim 23 is applied for, the biologic stabilizer is included. 27. The article of claim 26, wherein the stabilizer is an anticoagulant, a preservative, a protein plum suppressor, or a combination thereof. Hanru applied for the item of item 27, and the anticoagulation (4) was citrate, ethyldiaminetetraacetic acid, heparin, oxalate, vapor or any combination thereof. 29. If the patent application scope is 27 items of the Chinese brother, the reduction agent is sodium, sodium citrate and sodium borate. 30. The object of claim 27, wherein the protease inhibitor is dipeptide peptidase IV. 31. The object of claim 26, wherein the peptide and/or the stabilizer are donggan. 32. A composition comprising a sample derived from a mammalian body and a peptide as claimed in claim 1 of the 2008. 33. The composition of claim 32, further comprising a biological stabilizer. 34. The composition of claim 33, wherein the stabilizer is an anticoagulant, a preservative, a protease inhibitor, or a combination thereof. 35. The composition of claim 34, wherein the anticoagulant is citrate, ethyldiaminetetraacetic acid, heparin, oxalate, fluoride or a combination thereof. 36. The composition of claim 34, wherein the preservative is boric acid, sodium formate and sodium borate. 37. The composition of claim 34, wherein the protease inhibitor is dipeptide peptidase IV. 38. The composition of claim 32, wherein the sample is a blood product. 39. The composition of claim 38, wherein the blood product is blood plasma, platelets, white blood cells or stem cells. 40. A method of deactivating a virus comprising contacting the virus with a peptide as in item 1 of the patent application. 41. The method of claim 40, wherein the virus is a flavivirus, a hepatic virus or a human immunodeficiency virus. 42. A method of preventing or treating a mammalian cell infected with a virus comprising contacting the mammalian cell with a peptide as claimed in claim 1 of the scope of the patent. 43. The method of claim 42, wherein the virus is a flavivirus, liver 14 200837075 inflammatory virus or human immunodeficiency virus. For example, the method of claim 42 wherein the virus is a hepatitis C virus, West Nile virus, dengue virus or human immunodeficiency virus. 45. The method of claim 42, wherein the mammalian cell is a human cell. A method of preventing or treating a viral infection in a mammal, comprising administering to the mammal an effective amount of a peptide as in claim 1 of the scope of the patent application. • 47. A method of claiming the shame of the patent scope, wherein the mammal is a human. 48. The method of claim 46, wherein the virus is a flavivirus, a hepatic virus or a human immunodeficiency virus. The method of claim 46, wherein the virus is hepatitis C disease I, West Nile virus, dengue virus or human immunodeficiency virus. The method of claim 46, wherein the peptide is administered topically or systemically. , a method for deactivating a human immunodeficiency virus, comprising contacting the virus with a peptide of from 14 to 50 amino acids or L-amino acid, wherein the peptide comprises/spiral structure, and wherein The polar amino acid is on the same side of the alpha-helical structure and the non-polar amino acid is on the other side of the helical structure. 52. The method of claim 51, wherein the non-polar amino acid is selected from the group consisting of alanine, lysine, leucine, methionine, isoleucine, phenylalanine, and color a group consisting of amino acids. • The method of claim 51, wherein the polar amino acid is selected from the group consisting of arginine, aspartame, aspartic acid, cysteine, glutamic acid, bran 15 53 200837075 amine a group consisting of histidine, homocysteine, lysine, lysine, ornithine, serine and threonine. 54. The method of claim 53, wherein the cysteine is located at one of the N-terminus of the serine acid on the peptide. 55. As for the branch of item (4), the money is located at four positions of the N-terminus of the serine on the peptide. 56. If you apply for a special (4) 53-shaped branch, the cysteine is located at position 11 relative to the N-terminus of the peptide. 57. The method of applying for a patent of the patent, wherein the amino acid 16 and the amino acid 18 are charged with respect to the peptide, and wherein the amino acid and the amino acid are positively and negatively charged, Positive and positive, negative or positive. As described in the method of claim 5, the method 1 towel peptide is from M to 40 D-amino acids or l-amino acids. 59. For the branch of item 51, the (4) position is 14 to 3 D-amino acids or L-amino acids. 60. For example, the nodule of 51 items is rhyme to paste D-amino acid or l-amino acid. For example, it is difficult to apply (4) the branch of 51, the secret peptide is long post (8) solid D-amino acid or L-amino acid. 62. If applying for the method of item 61, the towel (4) has a peptide length of 14 amino groups ϊ夂 '(b) ehai and other amino acids are arginine, hemi-amic acid, valine, silk Amino acid, lysine, two aspartic acid, two leucine, two isoleucine, and three tryptophan residues; and (4) the oleic acid, semi-deaminic acid, face amine The acid, serine, and aspartate residues are on the same side of the helical structure of the core. The method of claim 62, wherein the amino acid sequence of the peptide is: SWLRDIWDWICEVL (SEQ ID NO: 92), or LVECIWDWIDRLWS (SEQ ID NO. 102). 64. The method of claim 61, wherein (4) the peptide is 15 amino acids long; (b) the amino acids are arginine, cysteine, glutamic acid, and two silk amines. Acid, proline, two aspartic acid, two leucine, two isoleucine, and three tryptophan residues; and (c) the arginine, cysteine, bran The amine acid, the serine acid, and the aspartic acid residue are on the same side of the helix structure. 65. The method of claim 64, wherein the amino acid sequence of the peptide is: SWLRDIWDWICEVLS (SEQ ID NO: 93), or SLVECIWDWIDRLWS (SEQ ID NO: 101). 66. The method of claim 61, wherein (4) the peptide is 16 amino acids in length; (b) the amino acids are arginine, cysteine, glutamic acid, and two silk amines. Acid, valine, three aspartic acid, two leucine, two isoleucine, and three tryptophan residues; and (c) the arginine, cysteine, bran The amine acid, the serine acid, and the aspartic acid residue are on the same side of the alpha-helical structure. 67. The method of claim 66, wherein the peptide amino acid sequence SWLRDIWDWICEVLSD (SEQ ID NO: 94), or DSLVECIWDWIDRLWS (SEQ ID NO: 100) 〇 17 200837075 68. The method of (a) the peptide is 17 amino acids long; (b) the amino acids are arginine, cysteine, face acid, two serines, valine, Three aspartic acid, two leucine, two isoleucine, three tryptophan and monophenylalanine; and (c) the arginine, cysteine, face acid, The serine and aspartic acid residues are located on the same side of the 〇: _ helical structure. 69. The method of claim 68, wherein the amino acid sequence of the peptide is: SWLRDIWDWICEVLSDF (SEQ ID NO: 95), or FDSLVECIWDWIDRLWS (SEQ ID NO: 99). 70. The method of claim 61, wherein (a) the peptide is 18 amino acids long; (b) the amino acids are arginine, cysteine, glutamic acid, two Serine, valine, three aspartic acid, two leucines, two isoleucines, three tryptophan acids, monophenylalanine and mono-amino acid; and (c) such The arginine, cysteine, glutamic acid, serine, and aspartic acid residues are on the same side of the alpha-helical structure. 71. The method of claim 70, wherein the amino acid sequence of the peptide is: SWLRDIWDWICEVLSDFK (SEQ ID NO: 43), KFDSLVECIWDWIDRLWS (SEQ ID NO: 96), SIWRDWVDLICEFLSDWK (SEQ ID NO: 97) Or KWLCRIWSWISDVLDDFE (SEQ ID NO: 98). 72. The method of claim 51, wherein the peptide EC5o is about 3 or less. The method of claim 51, wherein the peptide EC5〇 is about 2//Μ or less. 74. The method of claim 51, wherein the peptide EC5〇 is about 1//Μ or less. 75. The method of claim 51, wherein the peptide EC5〇 is about 500 nM or less. 76. The method of claim 51, wherein the peptide has an EC50 of about 400 nM or less. 77. The method of claim 51, wherein the peptide EC5〇 is about 300 nM or less. 78. The method of claim 51, wherein the peptide comprises an amino acid sequence selected from the group consisting of: SWLRPIWPWICEVLSDFK (SEQ ID NO: 91), SWLRDIWDWICEVL (SEQ ID NO: 92)? SWLRDIWDWICEVLS (SEQ ID NO: 93), SWLRDIWDWICEVLSD (SEQ ID NO: 94) 5 SWLRDIWDWICEVLSDF (SEQ ID NO: 95)? KFDSLVECIWDWIDRLWS (SEQ ID NO: SEQ ID NO: 95) : 96), SIWRDWVDLICEFLSDWK (SEQ ID NO: 97), KWLCRIWSWISDVLDDFE (SEQ ID NO: 98) 5 FDSLVECIWDWIDRLWS (SEQ ID NO: 99)? DSLVECIWDWIDRLWS (SEQ ID NO: 100)? SLVECIWDWIDRLWS (SEQ ID NO: 101), and LVECIWDWIDRLWS (SEQ ID NO: 102). The method of claim 51, wherein the peptide is composed of one amino acid sequence selected from the group consisting of SEQ ID NO: 91-102. 0 19 200837075 80. The method of claim 51, wherein the peptide comprises an amino acid sequence selected from the group consisting of: QIVGGVYLLPRRGPRLGV (SEQ ID NO: 4), QPGYPWPLYGNEGCGWAG (SEQ ID NO: 5), LYGNEGCGWAGWLLSPRG ( SEQ ID NO: 6), GWAGWLLSPRGSRPSWGP (SEQ ID NO: 7), IFLLALLSCLTVPASAYQ (SEQ ID NO: 8), DAILHTPGCVPCVREGNA (SEQ ID NO: 9), LPTTQLRRHIDLLVGSAT (SEQ ID NO: 10)? RHIDLLVGSATLCSALYV (SEQ ID NO: 11 ), GSATLCSALYVGDLCGSV (SEQ ID NO: 12), ALYVGDLCGSVFLVGQLF (SEQ ID NO: 13), IMDMIAGAHWGVLAGIAY (SEQ ID NO: 14), HINSTALNCNESLNTGWL (SEQ ID NO: 15), NCNESLNTGWLAGLFYQH (SEQ ID NO: 16), LASCRRLTDFAQ GWGPIS (SEQ ID NO: 17), TDFAQGWGPISYANGSGL (SEQ ID NO: 18), GPISYANGSGLDERPYCW (SEQ ID NO: 19), GSGLDERPYCWHYPPRPC (SEQ ID NO: 20)? WMNSTGFTKVCGAPPCVI (SEQ ID NO: 21), PCVIGGVGNNTLLCPTDC (SEQ ID NO: 2) 2), MYVGGVEHRLEAACNWTR (SEQ ID NO: 23), YLYGVGSSIASWAIKWEY (SEQ ID NO: 24), SIASWAIKWEYVVLLFLL (SEQ ID NO: 25), KWEYVVLLFLLLADARVC (SEQ ID NO: 26), WMMLLISQAEAALENLVI (SEQ ID NO: 27), GAVYAFYGMWPLLLLLLA ( SEQ ID NO: 28), GMWPLLLLLLALPQRAYA (SEQ ID NO: 29) 5 TLVFDITKLLLAIFGPLW (SEQ ID NO: 30), VSTATQTFLATCIN (SEQ ID NO: 31), ATQTFLATCINGVCWTVY (SEQ ID NO: 32), DSSVLCECYDAGCAWYEL (SEQ ID NO: 33 )? AYMNTPGLPVCQDHLEFW (SEQ ID NO: 34), LEFWEGVFTGLTHIDAHF (SEQ ID NO: 35), 20 200837075 HPITKYIMTCMSADLEW (SEQ ID NO: 36), VTSTWVLVGGVLAAL (SEQ ID NO: 37), WVLVGGVLAALAAYCLST (SEQ ID NO: 38), LAALAAYCLSTGCVV (SEQ ID NO. 39), EVFWAKHMWNFISGIQYL (SEQ ID NO: 40), MWNFISGIQYLAGLSTLP (SEQ ID NO) : 41), PAILSPGALVVGVVCAAI (SEQ ID NO: 42), SWLRDIWDWICEVLSDFK (SEQ ID NO: 43) 9 DWICEVLSDFKTWLKAKL (SEQ ID NO: 44), YVSGMTTDNLKCPCQIPS (SEQ ID NO: 45), SSGADTEDVVCCSMS (SEQ ID NO: 46), DTEDVVCCSMSYSW (SEQ ID NO: 47) 5 SSGADTEDVVCCSMSYSW (SEQ ID NO: 48)? DVVCCSMSYSWTGAL (SEQ ID NO: 49), TVTESDIRTEEAIYQCCD (SEQ ID NO: 50), GNTLTCYIKARAACRAAG (SEQ ID NO: 51), RAAGLQDCTMLVCGDDLV (SEQ ID NO: 52), CTMLVCGDDLVVICESAG (SEQ ID NO: 53), DDLVVICESAGVQEDAAS (SEQ ID NO: 54), LELITSCSSNVSVAHDGA (SEQ ID NO: 55), HTPVNSWLGNIIMFAPTL (SEQ ID NO: 56), APTLWARMILMTHFFSVL (SEQ ID NO: 57)? DQLEQALNCEIYGACYSI ( SEQ ID NO: 58), GVPPLRAWRHRARSVRAR (SEQ ID NO: 59), WRHRARSVRARLLSRGGR (SEQ ID NO: 60), GWFTAGYSGGDIYHSVSH (SEQ ID NO: 61), LYGNEGLGWAGWLLSPRG (SEQ ID NO: 62), IFLLALL SCITVPVSAAQ (SEQ ID NO: 63), IFLLALLSCLTIPASAYE (SEQ ID NO: 64), MSATFCSALYVGDLCGGV (SEQ ID NO: 65), GAAALCSAMYVGDLCGSV (SEQ ID NO: 66), ALYVGDLCGGVMLAAQVF (SEQ ID NO: 67), AMYVGDLCGSVFLVAQLF (SEQ ID NO) :68), IIDIVSGAHWGVMFGLAY (SEQ ID NO: 69) 5 VVDMVAGAHWGVLAGLAY (SEQ ID NO: 70), 21 200837075 VDVQYMYGLSPAITKYVV (SEQ ID NO: 71), YLYGIGSAVVSFAIKWEY (SEQ ID NO: 72)? WMLILLGQAEAALEKLVV (SEQ ID NO: 73) , WMMLLIAQAEAALENLVV (SEQ ID NO: 74), GVVFDITKWLLALLGPAY (SEQ ID NO: 75)? ELIFTITKILLAILGPLM (SEQ ID NO: 76), VSQSFLGTTISGVLWTVY (SEQ ID NO: 77), ATQSFLATCVNGVCWTVY (SEQ ID NO: 78), SWLRDVWDWVCTILTDFK (SEQ ID NO: 79), SWLRDVWDWICTVLTDFK (SEQ ID NO: 80) 5 DWVCTILTDFKNWLTSKL (SEQ ID NO: 81)? DWICTVLTDFKTWLQSKL (SEQ ID NO: 82), ASEDVYCCSMSYTWT (SEQ ID NO: 83), EDDTTVCCSMSYSW (SEQ ID NO: 84), CTMLVCGDDLVVICESAG (SEQ ID NO: 85), and PTMLVCG DDLVVISESQG (SEQ ID NO: 86). The method of claim 51, wherein the peptide consists of one amino acid sequence selected from the group consisting of SEQ ID NO: 4-86. 82. The method of claim 51, wherein the peptide comprises an amino acid sequence selected from the group consisting of: LYGNEGCGWAGWLLSPRG (SEQ ID NO: 6), IFLLALLSCLTVPASAYQ (SEQ ID NO: 8) ), GSATLCSALYVGDLCGSV (SEQ ID NO: 12), ALYVGDLCGSVFLVGQLF (SEQ ID NO: 13), IMDMIAGAHWGVLAGIAY (SEQ ID NO: 14), WMNSTGFTKVCGAPPCVI (SEQ ID NO: 21), MYVGGVEHRLEAACNWTR (SEQ ID NO: 23), YLYGVGSSIASWAIKWEY (SEQ ID NO: 24), WMMLLISQAEAALENLVI (SEQ ID NO: 27), GAVYAFYGMWPLLLLLLA (SEQ ID NO: 28), TLVFDITKLLLAIFGPLW (SEQ ID NO: 30), ATQTFLATCINGVCWTVY (SEQ ID NO: 32), 22 200837075 VTSTWVLVGGVLAAL (SEQ ID NO: 37), DWICEVLSDFKTWLKAKL (SEQ ID NO: 44) 5 DTEDVVCCSMSYSW (SEQ ID NO: 47), SSGADTEDVVCCSMSYSW (SEQ ID NO: 48), and CTMLVCGDDLVVICESAG (SEQ ID NO: 53). 83. The method of claim 51, wherein the peptide is selected from the group consisting of SEQ ID NO: 6, 8, 12, 13, 14, 2, 23, 24, 27, 28, 30, 32, Amino acid sequence consisting of one of the groups consisting of 37, 44, 47, 48 and 53. 84. The method of claim 51, wherein each of the amino acids in the indole peptide is a D-amino acid. 85. The method of claim 51, wherein each of the amino acids in the peptide is an L-amino acid. 86. The method of claim 51, wherein the peptide further comprises a dimethylaminonaphthalene moiety. 87. A method for preventing or treating a mammalian cell infected with a human immunodeficiency virus, comprising contacting the cell with a peptide of 14 to 5 amino acids or one amino acid, wherein the peptide comprises a peptide a ^-helical structure, and wherein the polar amino acid is on the same side of the alpha helix structure, and the non-polar amino acid is on the other side of the alpha helix structure. 88. The method of claim 87, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4-86 and 91-102. The method of claim 87, wherein the peptide consists of one amino acid sequence selected from the group consisting of SEQ ID NO. 4-86 and 91-102. 23 200837075 90. A method for preventing or treating a mammal infected with a human immunodeficiency virus, comprising administering to the mammal a long peptide of 14 to 50 D-amino acids or L-amino acids, wherein The peptide comprises an alpha-helical structure, and wherein the polar amino acid is on the same side of the alpha-helical structure and the non-polar amino acid is on the other side of the alpha-helical structure. 91. The method of claim 90, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4-86 and 91-102. 92. The method of claim 90, wherein the peptide consists of one amino acid sequence selected from the group consisting of SEQ ID NOs: 4-86 and 91-102. 93·- Isolated peptide of 14 to 50 D-amino acids or L-amino acids, comprising the amino acid sequence of formula V: XaarXaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9 -Xaa1〇-XaairW-Xaai3 W-X(10) 15_X%6-Xaair L_W_Xaa20_Xaa2rXaa22 (SEQ ID NO: 595), wherein Xaai is tryptophanic acid (w) or absent; Xaa2 is sulphate (τ) or is absent Xaas is lysine (Κ), glutamic acid (Ε), arginine (R), aspartic acid (D) or is absent; Xaa4 is phenylalanine or is absent; Xaa5 is ascending Acid (D), arginine (8), glutamic acid (E), lysine (6) or absent; Xaa0 is serine (s), sulphate (τ), leucine (L) or Does not exist. Xaa? is leucine (L) or lysine (v); 24 200837075 Xaas is proline (V), alanine (A) or serine; Xaap is glutamic acid (E), Amino acid (K), aspartic acid (D), threonine (τ), histidine (H), serine (S) or arginine (R); Xaa1G is cysteine (C) ), glutamic acid (E), leucine (L), serine (S) or arginine (R); Xaan is isoleucine (I) or proline (v); W is color Aminic acid; Xaan is aspartic acid (D), arginine (R) Gluten (6) or lysine (K); Xaa15 is isoleucine (I) or valine (V); Xaa! 6 is aspartic acid (D), arginine (R), glutamic acid (E), alanine (A), isoleucine (1) or lysine (K); Xaa17 is arginine (R), aspartic acid (D), lysine (K), tryptophan ( W) or glutamic acid (E); L is leucine; Xaa2G is serine (S), aspartic acid (D) or threonine (T); Xaa2i is glycine (G) or not Existence; and Xaa22 is serine acid (S) or absent; and wherein the amino acid sequence of formula V is not KFDSLVECIWDWIDRLWS (SEQ ID NO: 96), FDSLVECIWDWIDRLWS (SEQ ID NO: 99), DSLVECIWDWIDRLWS (SEQ ID NO: 100), SLVECIWDWIDRLWS (SEQ ID NO: 101), or LVECIWDWIDRLWS (SEQ ID NO: 102). 94. The peptide according to claim 93, which comprises an amino acid selected from the group consisting of: 25 200837075 KFDLVSECIWDWIDLRWS (SEQ ID NO: 277), FDLVSECIWDWIDLRWS (SEQ ID NO: 278) , DLVSECIWDWIDLRWS (SEQ ID NO: 279), LVSECIWDWIDLRWS (SEQ ID NO: 280), VSECIWDWIDLRWS (SEQ ID NO: 281), KFDSVVSCVWDWIIRLWD (SEQ ID NO: 282), FDSVVSCVWDWIIRLWD (SEQ ID NO: 283), DSVVSCVWDWIIRLWD (SEQ ID NO: 284)? SVVSCVWDWIIRLWD (SEQ ID NO: 285), VVSCVWDWIIRLWD (SEQ ID NO: 286), KFDSLVHLVWDWVEWLWS (SEQ ID NO: 288), FDSLVHLVWDWVEWLWS (SEQ ID NO: 289)? DSLVHLVWDWVEWLWS (SEQ ID NO: 290)? SLVHLVWDWVEWLWS (SEQ ID NO: 291), LVHLVWDWVEWLWS (SEQ ID NO: 292), KFDTLATCVWDWIARLWT (SEQ ID NO: 293), FDTLATCVWDWIARLWT (SEQ ID NO: 294), DTLATCVWDWIARLWT (SEQ ID NO: 295), TLATCVWDWIARLWT (SEQ ID NO) : 296), LATCVWDWIARLWT (SEQ ID NO: 297), KFDSLVTCVWDWVDRLWS (SEQ ID NO: 298), FDSLVTCVWDWVDRLWS (SEQ ID NO: 299)? DSLVTCVWDWVDRLWS (SEQ ID NO: 300), SLVTCVWDWVDRLWS (SEQ ID NO: 301), LVTCV WDWVDRLWS (SEQ ID NO: 302), KFDSLVESI WDWIDRLWS (SEQ ID NO: 303), FDSLVESI WDWIDRLWS (SEQ ID NO: 304), DSLVESI WDWIDRLWS (SEQ ID NO: 305), SLVESI WDWIDRLWS (SEQ ID NO: 306) 9 LVESIWDWIDRLWS (SEQ ID NO) : 307), EFRSLVKCIWRWIRDLWS (SEQ ID NO: 308), FRSLVKCIWRWIRDLWS (SEQ ID NO: 309)? RSLVKCIWRWIRDLWS (SEQ ID NO: 310) 5 SLVKCIWRWIRDLWS (SEQ ID NO: 311), LVKCIWRWIRDLWS (SEQ ID NO: 312), 26 200837075 EFDSLVECIWDWIDDLWS (SEQ ID NO: 313), FDSLVECIWDWIDDLWS (SEQ ID NO: 314), DSLVECIWDWIDDLWS (SEQ ID NO: 315), SLVECIWDWIDDLWS (SEQ ID NO: 316), LVECIWDWIDDLWS (SEQ ID NO: 317) 9 KFRSLVKCIWRWIRRLWS (SEQ ID NO: 318), FRSLVKCIWRWIRRLWS (SEQ ID NO) : 319), RSLVKCIWRWIRRLWS (SEQ ID NO: 320), SLVKCIWRWIRRLWS (SEQ ID NO: 321), LVKCIWRWIRRLWS (SEQ ID NO: 322), RFESLVDCIWEWIEKLWS (SEQ ID NO: 323)? FESLVDCIWEWIEKLWS (SEQ ID NO: 324), ESLVDCIWEWIEKLWS (SEQ ID NO: 325), SLVDCIWEWIEKLWS (SEQ ID NO: 326), LVDCIWEWIEKLWS (SEQ ID NO: 327), RFDSLVECIWDWIDKLWS (SEQ ID NO: 328), FDSLVECIWDWIDKLWS (SEQ ID NO: 329), DSLVECIWDWIDKLWS (SEQ ID NO: 330)? SLVECIWDWIDKLWS (SEQ ID NO: 331), LVECIWDWIDKLWS (SEQ ID NO: 332), KFDSLVECIWDWIDKLWS (SEQ ID NO: 333), FDSLVECIWDWIDKLWS (SEQ ID NO: 334)? DSLVECIWDWIDKLWS (SEQ ID NO: 335), SLVECIWDWIDKLWS ( SEQ ID NO: 336), LVECIWDWIDKLWS (SEQ ID NO: 337), KFESLVDCIWEWIERLWS (SEQ ID NO: 338), FESLVDCIWEWIERLWS (SEQ ID NO: 339), ESLVDCIWEWIERLWS (SEQ ID NO: 340), SLVDCIWEWIERLWS (SEQ ID NO: 341 ), LVDCIWEWIERLWS (SEQ ID NO: 342), KFESLVECIWEW1ERLWS (SEQ ID NO: 343), FESLVECIWEWIERLWS (SEQ ID NO: 344), ESLVECIWEWIERLWS (SEQ ID NO: 345), SLVECIWEWIERLWS (SEQ ID NO: 346), LVECIWEWIERLWS (SEQ ID NO) : 347), 27 200837075 EFRSLVKCIWRWIRDLWS (SEQ ID NO: 348), FRSLVKCIWRWIRDLWS (SEQ ID NO: 349), RSLVKCIWRWIRDLWS (SEQ ID NO: 350), SLVKCIWRWIRDLWS (SEQ ID NO: 351), LVKCIWRWIRDLWS (SEQ ID NO: 352) , EFDSLVECIWDWIDDLWS (SEQ ID NO: 353), FDSLVECIWDWIDDLWS (SEQ ID NO: 354), DSLVECIWDWIDDLWS (SEQ ID NO: 355), SLVECIWDWIDDLWS (SEQ ID NO: 356), LVECIWDWIDDLWS (SEQ ID NO: 357), KFRSLVKCIWRWIRRLWS (SEQ ID NO: 358)? FRSLVKCIWRWIRRLWS (SEQ ID NO: 359), RSLVKCIWRWIRRLWS (SEQ ID NO: 360), SLVKCIWRWIRRLWS (SEQ ID NO: 361), LVKCIWRWIRRLWS (SEQ ID NO: 362), KFDSLVERIWDWIDRLWS (SEQ ID NO: 363)? FDSLVERIWDWIDRLWS (SEQ ID NO: 364), DSLVERIWDWIDRLWS (SEQ ID NO: 365), SLVERIWDWIDRLWS (SEQ ID NO: 366), LVERIWDWIDRLWS (SEQ ID NO: 367), KFDSLVEEIWDWIDRLWS (SEQ ID NO: 368), FDSLVEEIWDWIDRLWS (SEQ ID NO: 369), DSLVEEIWDWIDRLWS (SEQ ID NO: 370), SLVEEIWDWIDRLWS (SEQ ID NO: 371), LVEEIWDWIDRLWS (SEQ ID NO: 372), KFDSLVECIWDWIDRLWSGS (SEQ ID NO: 373), FDSLVECIWDWIDRLWSGS (SEQ ID NO) : 374), DSLVECIWDWIDRLWSGS (SEQ ID NO: 375), SLVECIWDWIDRLWSGS (SEQ ID NO: 376), LVECIWDWIDRLWSGS (SEQ ID NO: 377), KFDSLVECIWDWIDRLWSG (SEQ ID NO: 378), FDSLVECIWDWIDRLWSG (SEQ ID NO: 379), DSLVECIWDWIDRLWSG (SEQ ID NO: 380), SLVECIWDWIDRLWSG (SEQ ID NO: 381), LVECIWDWIDRLWSG (SEQ ID NO: 382), 28 200837075 TKFDSLVECIWDWIDRLWS (SEQ ID NO: 383), KFDSLVECIWDWIDRLWS (SEQ ID NO: 384), FDSLVECIWDWIDRLWS (SEQ ID NO: 385), DSLVECIWDWIDRLWS (SEQ ID NO: 386), SLVECIWDWIDRLWS (SEQ ID NO: 387), WTKFDSLVECIWDWIDRLWS (SEQ ID NO: 388), ; TKFDSLVECIWDWIDRLWS (SEQ ID NO: 389) 5 KFDSLVECIWDWIDRLWS (SEQ ID NO: 390) 5 A FDSLVECIWDWIDRLWS (SEQ ID NO: 391), and • DSLVECIWDWIDRLWS (SEQ ID NO: 392). φ 95. The peptide according to claim 93, which consists of one amino acid sequence selected from the group consisting of SEQ ID NO: 277-392. 96. The peptide of claim 93, which comprises the amino acid sequence of formula VI: Xaai-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-LV-Xaa9-CIW-χ^13·\ν·ΚΧ^ 16-Χ&α17-Ι^\ν·8-Χ^21-Χαα22 (SEQ ID NO: 596) wherein: Xaa, Xaa2, Xaa21&Xaa22 are absent; 'Xaas is lysine (K), bran Amine acid (E), aspartic acid (D), arginine (8) Φ or is absent; > Xaa4 is phenylalanine (F) or is absent; Xaas is aspartic acid (D), spermine Acid (8), lysine (κ) or absent; Xaa0 is serine (S) or absent; L is leucine; V is valine; XaaP is lysine (K), spermine Acid (R), glutamic acid (E) or aspartic acid (D); C is cysteine; I is isoleucine; w is tryptophan; Xaan is aspartic acid (D), spermine Acid (R) or lysine (κ); Xaa! 6 is arginine (8), aspartic acid (d) or lysine (κ); 29 200837075 Xaa17 & arginine (R), aspartic acid ( D), glutamic acid (E) or lysine (K); and S is serine. 97. The peptide according to claim 96, which comprises an amino acid selected from the group consisting of: DFKSLVRCIWKWIKELWS (SEQ ID NO: 481); FKSLVRCIWKWIKELWS (SEQ ID NO: 482); KSLVRCIWKWIKELWS (SEQ ID NO: 483); SLVRCIWKWIKELWS (SEQ ID NO: 484); LVRCIWKWIKELWS (SEQ ID NO: 485); KFDSLVECIWKWIKRLWS (SEQ ID NO: 486); FDSLVECIWKWIKRLWS (SEQ ID NO: 487); DSLVECIWKWIKRLWS (SEQ ID NO: 488); SLVECIWKWIKRLWS (SEQ ID NO: 489); LVECIWKWIKRLWS (SEQ ID NO: 490); KFKSLVKCIWDWIDRLWS (SEQ ID NO: 491); FKSLVKCIWDWIDRLWS (SEQ ID NO: 492); KSLVKCIWDWIDRLWS (SEQ ID NO: 493); SLVKCIWDWIDRLWS ( SEQ ID NO: 494); LVKCIWDWIDRLWS (SEQ ID NO: 495); KFDSLVECIWRWIRRLWS (SEQ ID NO: 496); FDSLVECIWRWIRRLWS (SEQ ID NO: 497); DSLVECIWRWIRRLWS (SEQ ID NO: 498); SLVECIWRWIRRLWS (SEQ ID NO: 499 LVECIWRWIRRLWS (SEQ ID NO: 500); KFRSLVRCIWDWIDRLWS (SEQ ID NO: 501); FRSLVRCIWDWIDRLWS (SEQ ID NO: 502); RSLVRCIWDWIDRLWS (SEQ ID NO: 503); SLVRCIWDWIDRLWS (SEQ ID NO: 504); LVRCIWDWIDRLWS (SEQ) ID NO: 505); KFDSLVRCIWDWI RRLWS (SEQ ID NO: 506); FDSLVRCIWDWIRRLWS (SEQ ID NO: 507); DSLVRCIWDWIRRLWS (SEQ ID NO: 508); 30 200837075 SLVRCIWDWIRRLWS (SEQ ID NO: 509); LVRCIWDWIRRLWS (SEQ ID NO: 510); KFDSLVKCIWDWIKRLWS (SEQ ID NO: 511); FDSLVKCIWDWIKRLWS (SEQ ID NO: 512); DSLVKCIWDWIKRLWS (SEQ ID NO: 513); SLVKCIWDWIKRLWS (SEQ ID NO) : 514); LVKCIWDWIKRLWS (SEQ ID NO: 515); KFRSLVECIWDWIRRLWS (SEQ ID NO: 516); FRSLVEC1WDWIRRLWS (SEQ ID NO: 517); RSLVECIWDWIRRLWS (SEQ ID NO: 518); SLVECIWDWIRRLWS (SEQ ID NO: 519); LVECIWDWIRRLWS (SEQ ID NO: 520); KFKSLVECIWDWIKRLWS (SEQ ID NO: 521); FKSLVECIWDWIKRLWS (SEQ ID NO: 522); KSLVECIWDWIKRLWS (SEQ ID NO: 523); SLVECIWDWIKRLWS (SEQ ID NO: 524); LVECIWDWIKRLWS (SEQ ID NO: 525); KFDSLVRCIWRWIDRLWS (SEQ ID NO: 526); FDSLVRCIWRWIDRLWS (SEQ ID NO: 527); DSLVRCIWRWIDRLWS (SEQ ID NO: 528); SLVRCIWRWIDRLWS (SEQ ID NO: 529); LVRCIWRWIDRLWS (SEQ ID NO: 530); KFDSLVKCIWKWIDRLWS (SEQ ID NO: 531); FDSLVKCIWKWIDRLWS (SEQ ID NO) : 532); DSLVKCIWKWIDRLWS (SEQ ID NO: 533); SLVKCIWKWIDRLWS (SEQ ID NO: 534); LVKCIWKWIDRLWS (SEQ ID NO: 535); KFRSLVRCIWDWIRDLWS (SEQ ID NO: 536); FRSLVRCIWDWIRDLWS (SEQ ID NO: 537); RSLVRCIWDWIRDLWS (SEQ ID NO: 538); SLVRCIWDWIRDLWS (SEQ ID NO: 539); LVRCIWDWIRDLWS (SEQ ID NO: 540); KFKSLVKCIWDWIDRLWS (SEQ ID NO: 541); FKSLVKCIWDWIDRLWS (SEQ ID NO: 542); KSLVKCIWDWIDRLWS (SEQ ID NO: 543); 31 200837075 SLVKCIWDWIDRLWS (SEQ ID NO.. 544); and LVKCIWDWIDRLWS (SEQ ID NO.. 545). 98. The peptide according to claim 96, which consists of one amino acid sequence selected from the group consisting of seq ID NO: 481 to 545. 99. The isolated peptide of claim 96, which comprises an amino acid sequence of formula VII: Xaa1-Xaa2-Xaa3_Xaa4-Xaa5-Xaa6-L_V-Xaa9-C_I_W-Xaa13-WI-Xaa16- RLWS-Xaa21-Xaa22 (SEQ ID NO: 597), wherein: Xaa, Xaa2, Xaa21&Xaa22 are absent; Xaa3 is lysine (κ) or absent; Xaa4 is phenylalanine (f) or Is absent; Xaas is arginine (R) or lysine or is absent; Xaa6 is serine (S) or is absent; L is leucine; V is valine; Xaa; Amine acid (K), glutamic acid (E) or aspartic acid (D); C is cysteine; I is isoleucine; w is tryptophan; Xaau is arginine (R) or Amino acid (κ); Xaa^ is aspartic acid (D), arginine (R) or lysine (κ); and R is arginine; S is serine. 100. The peptide according to claim 99, which comprises an amino acid selected from the group consisting of: KFRSLVKCIWRWIDRLWS (SEQ ID NO: 546); FRSLVKCIWRWIDRLWS (SEQ ID NO: 547); 200837075 RSLVKCIWRWIDRLWS (SEQ ID NO: 548); SLVKCIWRWIDRLWS (SEQ ID NO: 549); LVKCIWRWIDRLWS (SEQ ID NO: 550); KFKSLVKCIWKWIDRLWS (SEQ ID NO: 551); FKSLVKCIWKWIDRLWS (SEQ ID NO: 552); KSLVKCIWKWIDRLWS (SEQ ID NO) : 553); SLVKCIWKWIDRLWS (SEQ ID NO: 554); LVKCIWKWIDRLWS (SEQ ID NO: 555); KFKSLVECIWKWIKRLWS (SEQ ID NO: 556); FKSLVECIWKWIKRLWS (SEQ ID NO: 557); KSLVECIWKWIKRLWS (SEQ ID NO: 558); SLVECIWKWIKRLWS (SEQ ID NO: 559); LVECIWKWIKRLWS (SEQ ID NO: 560); KFRSLVECIWRWIRRLWS (SEQ ID NO: 561); FRSLVECIWRWIRRLWS (SEQ ID NO: 562); RSLVECIWRWIRRLWS (SEQ ID NO: 563); SLVECIWRWIRRLWS (SEQ ID NO: 564); LVECIWRWIRRLWS (SEQ ID NO: 565); KFRSLVDCIWRWIRRLWS (SEQ ID NO: 566); FRSLVDCIWRWIRRLWS (SEQ ID NO: 567); RSLVDCIWRWIRRLWS (SEQ ID NO: 568); SLVDCIWRWIRRLWS (SEQ ID NO) : 569); and LVDCIWRWIRRLWS (SEQ ID NO: 570). 101. The peptide according to claim 99, which consists of one amino acid sequence selected from the group consisting of SEQ ID NO: 546 to 570. 102. The isolated peptide of claim 96, which comprises an amino acid sequence of formula VIII: Xaai -Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-LV-Xaa9TMCIW-Xaai3"VV- I-Xaai6-Xaai7-LWS-Xaa2i-Xaa22 (SEQ ID NO: 598), wherein: Xaai, Xaa2, Xaa2i and Xaa22 are absent; 33 200837075 Xaa3 is arginine (r), lysine (K) or Is absent; Xaa4 is phenylalanine (F) or is absent; Xaas is arginine (R), lysine (K) or is absent; Xaa6 is serine (S) or is absent L is leucine; V is valine; Xaa9, Xaa13, 3 & 16 and 3317 are arginine (R) or lysine (κ); and C is cysteine; I is different Leucine; w is tryptophan; S is serine. 103. The peptide according to claim 102, which comprises an amino acid selected from the group consisting of KFKSLVKCIWKWIKRLWS (SEQ ID NO: 571); FKSLVKCIWKWIKRLWS (SEQ ID NO: 572); KSLVKCIWKWIKRLWS (SEQ ID NO: 573); SLVKCIWKWIKRLWS (SEQ ID NO: 574); LVKCIWKWIKRLWS (SEQ ID NO: 575); KFRSLVRCIWRWIRRLWS (SEQ ID NO: 576); FRSLVRCIWRWIRRLWS (SEQ ID NO: 577); RSLVRCIWRWIRRLWS (SEQ ID NO: 578); SLVRCIWRWIRRLWS (SEQ ID NO: 579); LVRCIWRWIRRLWS (SEQ ID NO: 580); RFRSLVRCIWRWIRRLWS (SEQ ID NO: 581); FRSLVRCIWRWIRRLWS (SEQ ID NO: 582); RSLVRCIWRWIRRLWS (SEQ ID NO: 583); SLVRCIWRWIRRLWS ( SEQ ID NO: 584); LVRCIWRWIRRLWS (SEQ ID NO: 585); KFKSLVKCIWKWIKKLWS (SEQ ID NO: 586); FKSLVKCIWKWIKKLWS (SEQ ID NO: 587); KSLVKCIWKWIKKLWS (SEQ ID NO: 588); SLVKCIWKWIKRLWS (SEQ ID NO: 589 ); and LVKCIWKWIKKLWS (SEQ ID NO: 590). 34. The peptide of claim 102, which consists of one amino acid sequence selected from the group consisting of SEQ ID NO: 571-590. 35